QuantaSmart™ For The
TriCarb
Reference Manual
®
Liquid Scintillation
Analyzer
Release History
Kit Number/Part Number Release Publication Date
7000052/1694267 A May 2004
Any comments about the documentation for this product should be addressed to:
User Assistance
PerkinElmer, Inc.
2200 Warrenville Road
Downers Grove, Illinois 60515
USA
Or emailed to:
info@perkinelmer.com
Notices
The information contained in this document is subject to change without notice. Except as specifically set
forth in its terms and conditions of sale, PerkinElmer makes no warranty of any kind with regard
to this document, including, but not limited to, the implied warranties of merchantability and fitness for a particular purpose. PerkinElmer shall not be liable for errors contained herein for incidental con-
sequential damages in connection with furnishing, performance or use of this material.
Copyright Information
This document contains proprietary information that is protected by copyright. All rights are reserved. No part
of this publication may be reproduced in any form whatso ever or translated into any language without the prior,
written permission of PerkinElmer, Inc.
Copyright © 2004 PerkinElmer, Inc.
Produced in the USA.
Trademarks
Registered names, trademarks, etc. used in this document, even when not specifically marked as such, are
protected by law.
PerkinElmer is a registered trademark of PerkinElmer, Inc.
IPA™ is a trademark of PerkinElmer, Inc.
QuantaSmart™ is a trademark of PerkinElmer, Inc.
Replay™ is a trademark of PerkinElmer, Inc.
SpectraView™ is a trademark of PerkineElmer, Inc.
TriCarb
® is a registered trademark of PerkinElmer, Inc.
Varisette™ is a trademark of PerkinElmer, Inc.
Microsoft
® is a registered trademark of Microsoft Corporation.
MS-DOS is a registered trademark of Microsoft Corporation.
Pentium
Teflon
Windows
WindowsXP
® is a registered trademark of Intel Corporation.
® is a registered trademark of E.I. DuPont Company.
® is a registered trademark of Microsoft Corporation.
® is a registered trademark of Microsoft Corporation.
Safety
Electrical Safety
Use proper plugs and good earth ground connections.
WARNING
For systems operating at voltages other then 115 volts AC or 220 volts AC, a
locally approved 3-prong plug may be required to correctly power the system.
WARNING
CAUTION: Do not move the fully assembled unit. Use both hands when lifting or moving any part
of the system. Carry each part from the bottom.
Wiring Specifications
Live (L) Brown lead
Neutral (N) Blue lead
Earth (E) Green/yellow lead
Cleaning the System
Clean the outer surfaces of the system by wiping them with a damp cloth and common laboratory
cleaner.
System Ventilation
For adequate ventilation of this equipment, a distance of 15cm must be kept from this unit and any
other surfaces.
Explanation of Symbols
You may find one or more of the following symbols used on labels on your system.
Symbol Explanation L’explication Erklarung
Alternating Current Courant alternatift Wechselstrom
Protective Earth Ground Mise a la terre Schutz -Erdun
On (Supply) Marche (alimentation) Ein
Off (Supply) Arret (alimentation) Aus
Caution-Attention: Risk of
Electrical Shock
Caution (refer to
accompanying
documents)
Serial Out Sortie serie Serieller Ausgang
Printer Imprimante Drucker
Monitor Moniteur Bildschirm
Fuse Label 1 Current
Warning
Attention Risque de choc Vorsicht Spannug
Gefahrliche
Attention: Voir les
documents cijoints
Etiquette d'advertissement
relatif au type et au
courant du fusible
Vorsicht Siehe
Begleitinformation
Sicherung/Stromstarke
Table of Contents
Chapter 1
Specifications. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
System Control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
Energy Range . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
Efficiency . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
Chi-Square . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
Physical Dimensions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
Detectors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
Environmental Requirements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
Varisette™ Sample Changer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
Electrical Requirements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
External Standard . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
Observed Background . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
Sample Cassettes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
Shielding . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
Sample Vials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
Chapter 2
How To.... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
How to Get Started . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
How to Perform the SNC (Self-Normalization and Calibration) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
Steps in Performing an Assay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
How to Perform an Assay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
How to Create a Password . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
How to Associate an Assay to a Protocol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
How to Count Samples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
How to Disassociate an Assay from a Protocol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
How to Edit an Assay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21
How to Print an Assay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
How to Manually Print a Report . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
How to Link a Quench Set to a Sample Nuclide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
How To Set Up a Quench Standards Assay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
How To Run an Alpha Beta Assay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
How To Run an Alpha Beta Standards Assay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28
How to Use the Replay Feature . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
How to Calculate Radioactive Decay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
How to Change the Time and Date . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32
i
Chapter 3
The System Computer. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
Attaching To A Network . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
Chapter 4
The System Software. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
Software Security . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
Main Window . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36
Output Window . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
The SpectraView Window . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38
The Instrument Status Bar . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39
The Protocols Tree . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40
The Replay Tree . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43
Menus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
File Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
Run Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48
View Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50
Libraries Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51
Tools Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55
IPA Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 58
Diagnostics Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59
Window Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60
Help Menu . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61
Spectral Displays . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62
Spectral Mapping . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62
Spectrum Unfolding . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63
The SpectraView Window . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64
Reports . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65
Printed Reports . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65
Electronic Reports . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66
ii
Chapter 5
Assays . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .67
CPM Assays . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67
DPM Assays . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68
FS DPM Assay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69
Direct DPM Assays . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71
Alpha Beta Assays . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72
Alpha Beta Standards Assay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75
Quench Standards Assay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 76
SPC Assay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 77
Defining an Assay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 78
Assay Parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 79
Count Conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 81
Count Corrections . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 87
Report Definition . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 90
Report Output . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 98
Special Files . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 102
Worklist . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 105
Chapter 6
Libraries . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 109
Sample Nuclides Library . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 110
Quench Standards Library . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 113
Alpha Beta Nuclide Library . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 114
Alpha Beta Standards Library . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 116
Chapter 7
Calibration and Normalization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121
Calibration/Normalization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121
IPA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 121
When to Perform these Procedures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 122
Instrument Performance Assessment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123
IPA Definition . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 123
IPA Charts & Tables . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 127
IPA Reports . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 130
SNC for an Instrument without Super Low Level Counting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131
SNC for Super Low Level Counting (with BGO detector guard) . . . . . . . . . . . . . . . . . . . . . . . . . . . 133
iii
Chapter 8
Advanced Features . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 135
Priostat . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 135
Group Priostat . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 135
Sample Priostat . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 136
The Replay Feature . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 152
Replay Output Window . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 155
High Sensitivity and Low Level Counting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 156
Super Low Level Counting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 157
Alpha Beta Counting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 159
Tandem Processing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 161
Chapter 9
Maintenance and Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 163
Preventative Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 163
Storing Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 164
Operational Errors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 165
IPA Errors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 167
Warnings and Messages . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 169
System Messages . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 169
Chapter 10
Calculations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 183
Background Correction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 183
Background Threshold . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 183
Chi-Square Calculation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 184
DPM (Disintegrations Per Minute) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 184
Efficiency . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 184
Figure of Merit Calculation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 184
Half-life Correction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 185
IPA Background . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 185
LCR (Low Count Reject) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 185
LUM (% Luminescence) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 185
Radioactivity Units . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 186
2 Sigma % (%2s) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 186
Radionuclide Decay Calculator . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 187
% Reference (% Ref) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 187
Chapter 11
Glossary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 189
iv
Chapter 12
Theory. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 197
Low Level Counting Theory . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 197
Alpha Beta Counting Theory . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 207
v
vi
SPECIFICATIONS
Chapter 1
Specifications
All instrument specifications are developed using sources whose activity is
referenced to NIST source activity and PerkinElmer Life and Analytical Sciences
reference methods.
System Control
The system is controlled by an IBM-compatible Pentium computer. The standard
computer is configured with one floppy disk drive, a CD-drive and one hard drive.
The computer contains a serial RS-232 communications port, a parallel printer
port, and a graphics adapter for the color monitor. All RS-232 communication
occurs through communications port one. Communications port two is used for
proprietary purposes and cannot be used in conjunction with any peripheral
devices.
Note: The system can be attached to a network using an Ethernet
Adapter kit option. For further information on this kit, contact yo ur
PerkinElmer representative.
Energy Range
This table represents the preset regions for the following nuclides:
Figure 1-1 Energy Range
You can define preset regions for any nuclide in the Sample Nuclides Library.
PerkinElmer Life and Analytical Sciences 1
CHAPTER 1
Efficiency
For Tritium in the range 0-18.6 keV, the minimum acceptable efficiency is 60%
(58% for 3170TR/SL). For Carbon-14 in the range 0-156keV, the minimum
acceptable efficiency is 95% (94% for 3170TR/SL). These values were generated
by PerkinElmer Life and Analytical Sciences at our Downers Grove, Illinois facility.
The exact values obtained at other instrument locations may vary. Counting
Efficiency is a parameter measured as part of Instrument Performance
Assessment (IPA). IPA is an option on the TriCarb® 2800TR and 2900TR.
Chi-Square
Chi-Square acceptable range: 7.63 to 36.19, performed as 20, 0.5 minute repeat
counts and tested at the 99% confidence limits. Chi-Square is a parameter
measured as part of Instrument Performance Assessment (IP A). IP A is an option
on the Tri-Carb 2900TR
Physical Dimensions
The following table displays the physical dimensions of all TriCarb models:
Figure 1-2 Physical Dimensions
Detectors
Two diametrically opposed high performance photomultiplier tubes (PMTs) are
coupled to a light-tight, reflective optical chamber. The detectors are located
below the sample changer at the rear of the instrument.
2 PerkinElmer Life and Analytical Sciences
SPECIFICATIONS
Environmental Requirements
The recommended operating ambient temperatures are 59°F to 95°F (15°C to
35°C).
The recommended operating relative humidity is 30% to 85% non-condensing
humidity (the system is for indoor use only).
The Pollution Category is number two.
The instrument should be located away from all sources of radiation (X-ray
equipment, radiation storage vaults, etc.). The instrument should be positioned
such that direct sunlight or unscreened fluorescent light will not enter the sample
changer. Direct light may affect the optical sensors within the sample changer,
causing erratic operation. Direct light may also induce fluorescence within your
samples, causing erratic results.
The system must be connected to a stable power source, through the use of
proper plugs and good earth grounding. The system provides automatic recovery
after a power failure. Power to the temperature control unit should not be
supplied through a power strip.
Varisette™ Sample Changer (optional on 2800TR, 2900TR)
The sample changer automatically moves the samples into position and loads
them down into the detector for counting. After the count is completed, the
sample is automatically unloaded and the next sample is loaded.
The sample changer typically moves the cassettes in a forward (counterclockwise)
direction using a pair of synchronously driven transport belts. If necessary, these
belts can run in the reverse (clockwise) direction (such as during recovery from a
power failure).
The instrument can hold up to:
408 large vials
720 small vials
720 4ml vials
PerkinElmer Life and Analytical Sciences 3
CHAPTER 1
Electrical Requirements
The following table represents the electrical requirements for all Tricarb models.
Figure 1-3 Electrical Requirements
External Standard
133
Ba, nominal less than 20µCi (for all models except the TriCarb 3170TR/SL).
133
Ba, nominal less than 1µCi (for the TriCarb 3170TR/SL).
Observed Background
Average values for Normal Count Mode:
Tritium:17.3 CPM
Carbon-14:24.3 CPM
These values were generated by PerkinElmer Life and Analytical Sciences at our
Downers Grove, Illinois facility. The exact values obtained at other instrument
locations may vary.
4 PerkinElmer Life and Analytical Sciences
SPECIFICATIONS
Sample Cassettes
Cassettes are the plastic racks which hold sample vials and allow them to be
moved on the sample changer deck. Samples are placed into cassettes that
accommodate either standard or small vials without adapters (4ml vials require
cassettes with adapters). The standard vial cassette can accommodate up to 12,
15-20ml vials and the mini-vial cassettes up to 18, 6-7ml mini-vials. A protocol
flag, which you attach to the cassette, identifies the protocol and assay conditions
that you have defined for use with your samples. The individual cassettes are
identified by unique numbers (cassette IDs) located at the end of each cassette.
The protocol flag and cassette ID are automatically read by the instrument to
provide Positive Sample Identification (PID) when using Worklists.
Figure 1-4 Sample Cassette
Shielding
The detector assembly is surrounded by a minimum of 2 inches of lead.
PerkinElmer Life and Analytical Sciences 5
CHAPTER 1
Sample Vials
Vials and caps must conform to the following sizes. Caps must not exceed the
diameter of the vial.
Figure 1-5 Vial Sizes
6 PerkinElmer Life and Analytical Sciences
HOW TO...
Chapter 2
How To...
This chapter presents a quick overview of common tasks performed with the
QuantaSmart™ software.
How to Get Started
When you are ready to begin a counting procedure, you will need to perform the
following tasks:
1. Calibrate and Normalize the instrument.
2. Select an assay type: Alpha Beta, Alpha Beta Standards, CPM Assay, DPM
Single, DPM Dual, DPM Triple, FS DPM, Direct DPM, Quench Standards, Single
Photon Counting. Note: DPM Dual is optional on 2800 series instruments. DPM
Triple is optional on 2800 and 2900 series instruments.
3. For any DPM assay except Direct DPM, create quench data.
4. Define and save the new assay parameters.
5. Associate (link) the assay parameters to a protocol.
6. Attach the correct protocol flag to the first casset te to be counted and load the
cassette(s) with samples.
7 . Begin sample counting. Do not use the system’s CD writer while the instrument
is counting.
PerkinElmer Life and Analytical Sciences 7
CHAPTER 2
How to Perform the SNC (Self-Normalization and Calibration)
SNC for an Instrument without Super Low Level Counting Ability
1. Define the IPA parameters in the IPA Definition window.
Figure 2-1 IPA Definition Window.
2. Reset the SNC protocol flag to the reset position (the flag is all the way to the
left when the plug is on the left end of the cassette).
3. Load the purged, unquenched Carbon-14 standard into the first position of the
cassette (this is at the same end as the protocol plug).
Caution: Do not use the unpurged, Low Level standards to
calibrate the instrument, even if the instrument is to be used in
Low Level, High Sensitivity or Super Low Level count mode.
8 PerkinElmer Life and Analytical Sciences
HOW TO...
4. Load the purged, unquenched Tritium standard into the second cassette
position.
5. Load the purged background standard into the third cassette position.
6. Load the instrument with cassettes.
7. Press the green flag start button to begin counting. Do not use the
system’s CD writer while the instrument is counting.
SNC for an Instrument with Super Low Level Counting Ability (with
BGO Detector Guard)
1. Define the IPA parameters in the IPA Definition window.
2. Reset the SNC protocol flag to the reset position (the flag is all the way to the
left when the plug is on the left end of the cassette).
3. Load the purged, unquenched Carbon-14 standard into the first position of the
cassette (this is at the same end as the protocol flag).
4. Load an empty vial into the second cassette position.
5. Load the purged, unquenched Tritium standard into the third cassette position.
6. Load the purged background standard into the fourth cassette position.
7. Load the instrument with cassettes.
8. Press the green flag start button to begin counting. Do not use the
system’s CD writer while the instrument is counting.
PerkinElmer Life and Analytical Sciences 9
CHAPTER 2
Steps in Performing an Assay
1. Select the File-Open Assay menu option. The Open Assa y window is displayed.
Figure 2-2 Open Assay Window.
10 PerkinElmer Life and Analytical Sciences
HOW TO...
2. In the Open Assay window, select the assay for which you would like to create
a password. Click the Open button. The Assay Definition window is displayed.
Figure 2-3 Assay Definition Window.
3. In the Assay Parameters tab, mark the Lock Assay box. The Password field
becomes enabled.
4. Enter a descriptive password in the Password field.
5. When you hav e finished editing, save the assay. The password must be used to
edit the assay after it is saved.
6. In the Sample Nuclides Library, define an appropriate nuclide for the assay, if
one does not already exist.
PerkinElmer Life and Analytical Sciences 11
CHAPTER 2
How to Perform an Assay
1. Select the File-New Assay menu option.
2. Select an assay type in the Select Assay Type window.
Figure 2-4 Select Assay Type Window.
3. In each of the seven tabs of the Assay Definition window, define the individual
assay parameters, as needed.
4. Save the assay using a descriptive name.
5. In the Protocols Tree of the main window, select the protocol to which you
would like to associate an assay.
6. Right click on that protocol flag number.
7. From the menu that is displayed, select Associate Assay. The Associate Assay
window is displayed.
12 PerkinElmer Life and Analytical Sciences
HOW TO...
Note:Y ou can also associate an assay to a flag using the File menu.
Figure 2-5 Protocol Tree.
Figure 2-6 Associate Assay Window.
PerkinElmer Life and Analytical Sciences 13
CHAPTER 2
8. Select the Assay that you would like to associate to a protocol flag number.
Click the Open button. The Data Paths window is displayed.
Figure 2-7 Data Paths Window.
9. In the Data Paths window, enter a User ID for the assay.
10. Enter an Additional Header, if necessary.
11. Enter an Output Data Path, if you would like the data to be saved in a directory
other than the default.
In the following example, the default path is:
C:\Packard\TriCarb\Results\2800TR\14c_cpm
12. Attach the appropriate protocol flag in reset position.
13. Load the cassette(s) with samples.
14. Load the instrument with cassettes.
15. Press the (green start flag) button to begin counting. Do not use the
system’s CD writer while the instrument is counting.
14 PerkinElmer Life and Analytical Sciences
HOW TO...
How to Create a Password
1. Select the File-New Assay menu option. The Select Assay Type window is
displayed.
Figure 2-8 Select Assay Type Window.
2. In the Select Assay Type window, select the type of assay you would like to
PerkinElmer Life and Analytical Sciences 15
CHAPTER 2
create. The Assay Definition window is displayed.
Figure 2-9 Assay Definition Window.
3. In the Assay Parameters tab, mark the Lock Assay box. The Password field
becomes enabled.
4. Enter a descriptive password in the Password field.
5. When you have completed the assay definition process, save the assay. The
password must be used to edit the assay after it is saved.
16 PerkinElmer Life and Analytical Sciences
HOW TO...
How to Associate an Assay to a Protocol
1. Select the protocol to which you would like to associate an assay.
Figure 2-10 Protocol Tree.
2. Right click on that protocol flag number.
3. From the menu that is displayed, select Associate Assay. The Associate Assay
window is displayed.
4. Select the Assay that you would like to associate to a protocol flag number.
Click the Open button. The Data Paths window is displayed.
5. In the Data Paths window, enter a User ID for the assay.
6. Enter an Additional Header, if necessary.
PerkinElmer Life and Analytical Sciences 17
CHAPTER 2
7. Enter an Output Data Path, if you are generating either an RTF or Delimited
Text file for the assay and would like the data to be saved in a directory other
than the default. Check the Use Default Output Data P ath bo x if you would lik e
the data to be saved in the default directory.
Figure 2-11 Associate Assay Window.
Figure 2-12 Data Paths Window.
18 PerkinElmer Life and Analytical Sciences
HOW TO...
How to Count Samples
1. Attach the protocol flag with the number matching the protocol flag to which
you have associated the assay.
2. Reset the protocol flag by moving the plastic arm all the way to the left. The
flag is all the way to the left when the flag is on the left end of the cassette.
Figure 2-13 Cassette.
3. Place all of the background, reference and sample vials into the appropriate
cassette positions.
4. Place the cassette(s) in the sample changer deck so that the protocol number
on the flag is facing you. The cassette should be positioned on the right side
along the far wall of the sample changer deck or after the last set of cassettes
already on the sample changer deck if the machine is in use.
5. Press the button to begin counting. Note: Do not use the system’s CD
writer while the instrument is counting.
PerkinElmer Life and Analytical Sciences 19
CHAPTER 2
How to Disassociate an Assay from a Protocol
1. In the Protocols Tree of the main window, select the assay you would like to
disassociate from a protocol number.
Figure 2-14 Protocol Tree.
2. Right click on that assay.
Note:You can also disassociate an assay from a flag using the File
menu.
3. From the menu that is displayed, select Disassociate Assay.
20 PerkinElmer Life and Analytical Sciences
HOW TO...
How to Edit an Assay
1. Select the File-Open Assay menu option. The Open Assa y window is displayed.
Figure 2-15 Open Assay Window.
PerkinElmer Life and Analytical Sciences 21
CHAPTER 2
2. In the Open Assay window, select the assay you would like to edit. Click the
Open button. The Assay Definition window is displayed.
Figure 2-16 Assay Definition Window.
3. In each of the seven tabs of the Assay Definition window, change the assay
parameters, as needed.
4. If you would like to save the assa y with the same name, click the OK button.
5. If you would like to save the assay using a different name, click the Save As
button. The Save As window is displayed.
6. Enter an appropriate name for the assay and click the Save button.
22 PerkinElmer Life and Analytical Sciences
HOW TO...
How to Print an Assay
1. Select the File-Print Assays Main menu option. The Select Assays to Print
window is displayed.
Figure 2-17 Select Assays to Print Window.
2. Select the assay you would like to print and click the Open button. The Print
window is displayed.
3. In the print window, select the appropriate print parameters and click the OK
button. A list of the Assay Parameters, Count Conditions, Count Corrections
and Reports defined for the assay will print.
How to Manually Print a Report
1. In the Protocols Tree of the main window, select the report icon ( ) that you
would like to print. The Output window is displayed.
2. In the Output window, select the print icon( ). The report of interest will
print once the icon is selected.
PerkinElmer Life and Analytical Sciences 23
CHAPTER 2
How to Link a Quench Set to a Sample Nuclide
Note:Refer to page 26 on how to set up a quench set standards
assay. This requires the single/dual/color DPM option on the
2800TR.
1. Select the Libraries-Sample Nuclides Main menu item. The Sample Nuclides
window is displayed.
Figure 2-18 Sample Nuclides Window.
24 PerkinElmer Life and Analytical Sciences
HOW TO...
2. Click one of the Quench Se t buttons. The Quench Standards window is
displayed.
Figure 2-19 Quench Standards Window.
3. Select the Quench Set Low if you are counting one nuclide in one counting
region, the Quench Set Medium if you are counting a second radionuclide in a
separate region and the Quench Set High if you are counting a third
radionuclide in a third region.
4. Select the name of the quench set you would like to link to the sample nuclide.
5. Click OK. The name of the quench set(s) you selected should appear in the
Sample Nuclides Library window on the Quench Set buttons.
PerkinElmer Life and Analytical Sciences 25
CHAPTER 2
How To Set Up a Quench Standards Assay
The following tasks are required when performing a Quench Standards Assay.
This requires the single/dual/color DPM option on the 2800TR.
1. Calibrate the instrument, if necessary.
2. Create a new assay, choosing Quench Standards as the assay type and define
the new assay parameters.
Or
Open an existing Quench Standards Assay and edit or review if necessary.
3. Save the assay in the Assays folder of the Packard\TriCarb directory.
4. Associate (link) the assay parameters with a protocol and attach the
corresponding protocol flag to the first cassette to be counted.
5. Load the cassette(s) with vials and load the instrument with cassettes.
6. Start the instrument.
Note:When using the Low Level count mode (if available), you
must NOT use quench standards which have been purged free of
oxygen with an inert gas. The oxygen quenching in unpurged
standards facilitates discrimination between background and true
beta events. Unpurged standards are available from PerkinElmer
Life and Analytical Sciences.
26 PerkinElmer Life and Analytical Sciences
HOW TO...
How To Run an Alpha Beta Assay not available on the 2800TR; optional on the
2900TR and 3100TR series
The following tasks are required when performing an Alpha Beta Assay.
1. Calibrate the Instrument, if necessary.
2. Define and run an Alpha Beta Standards Assay (page 28), if necessary, to
establish the optimal pulse decay discriminator value.
3. Define an alpha beta nuclide in the Alpha Beta Nuclide Library. Choose the
Standards Set from the Alpha Beta Standards Library to use the discriminator
setting from the Standard Set.
4. Create a new assay by choosing Alpha Beta as the assay type. On the Count
Conditions tab, select the desired alpha beta radionuclide name from the Alpha
Beta Nuclide Library. Define the remaining new assay parameters.
Or
Open an existing assay and edit or review if necessary .
5. Save the assay in the Assays folder of the Packard\TriCarb directory.
6. Associate (link) the assay parameters with a protocol number in the protocol
tree and attach the corresponding protocol clip to the first cassette to be
counted.
7. Load the cassette(s) with vials and load the instrument with cassettes.
8. Click the green start button at the top of the main window.
9. Start the instrument.
PerkinElmer Life and Analytical Sciences 27
CHAPTER 2
How To Run an Alpha Beta Standards Assay not available on the 2800TR;
optional on the 2900TR and 3100TR series
The following tasks are required to run the Alpha Beta Standards Assay. Two
standards are required, a pure beta emitter and a pure alpha emitter.
1. Choose the Alpha Beta Standards selection from the Libraries menu.
2. Click the Add button and enter the name for a new Alpha Beta Standard Set.
3. Choose Automatic for the Discriminator T ype if your pure alpha and pure beta
standards have an activity of at least 50,000 CPM each. If the activity in either
standard is less than 50,000 CPM, choose Manual as the Discriminator Type
by clicking on it.
4. The remaining fields are for inf ormation only. They are either default values or
values computed by the instrument.
5. Choose File-New and select Alpha Beta Standards as the assay choice from
the drop down menu.
OR
Open an existing assay and edit or review if necessary .
6. Click the Name button on the Count Conditions Tab to choose from the list of
Alpha Beta Standard names in the Alpha Beta Standards Library. Define the
other available parameters as desired.
Note:High Sensitivity or Low Level Count mode are not available
when counting Alpha Beta Standards, but will be available when
counting samples in an Alpha Beta assay
7. When the assay definition is complete, name and save the assay.
8. Associate (link) the assay parameters with an available protocol flag in the
protocol tree. Place the numbered protocol clip on a cassette.
9. Place the pure beta emitter standard in cassette position 1 and the pure alpha
emitter standard in cassette position 2.
10. Load the cassette onto the instrument and click the green start button at the
top of the main window.
11. After counting is complete, the misclassification (or spillover) curve and the
optimum discriminator value will be stored for the Standard Set. Review the
curve and discriminator setting, if desired, from the Alpha Beta Standards
Library.
28 PerkinElmer Life and Analytical Sciences
HOW TO...
How to Use the Replay Feature (optional on TriCarb 2800, standard on 2900/
3100 series)
1. In the main window, click on the Replay™ tab.
2. Select the results file that you would lik e to analyze. A listing of the results files
is displayed in the Replay Tree using file names with the following syntax:
User id \ assay_name \
yyyymmdd_militarytime
3. Right click on the selected file name.
4. Select Open for Replay. The Replay window is displayed.
PerkinElmer Life and Analytical Sciences 29
CHAPTER 2
Note:Use the View menu and select Refresh to verify that you have
the latest view of the Replay files.
Figure 2-20 Replay Conditions Window.
5. In the Replay Conditions tab, define the parameters, as needed for reanalysis
of sample data.
6. In the Report Definition tab, define any printed or electronic reports you would
like to generate for the reanalysis of sample data.
7. Click the Replay button. Any reports that you defined are generated after the
reanalysis of data occurs.
Note: Data processed with Replay does not change the original
data. All Replay changes are temporary.
30 PerkinElmer Life and Analytical Sciences
HOW TO...
How to Calculate Radioactive Decay
1. Select the Tools-Nuclide Decay menu option. The Radionuclide Decay window
is displayed.
Figure 2-21 Radionuclide Decay Window.
2. From the drop-down list in the Nuclide field, select the nuclide of interest if it
appears in the list; select manual if you would like to manually enter the
nuclide half-life information.
3. Enter Reference Activity, Date and Time.
4. Click the Start Decay Button.
5. The current DPM for the nuclide is displayed in the Current DPM Activity field.
PerkinElmer Life and Analytical Sciences 31
CHAPTER 2
How to Change the Time and Date
Double click on the date and time on the task bar. The Date/Time Properties
window will display. Use this window to change the date and time.
Note:If an error message displays stating that the user has
insufficient rights to change the date and time, follow the
procedure below.
1. On the Windows NT screen, go to Start, Shutdown, click on Restart the
computer?, and then click on Yes.
2. When the Windows NT logo displays, press Shift steadily until the Logon
Information window displays. Type
Password: field blank. Click OK.
The system will start NT and you will be logged on as Administrator.
3. On the Windows NT screen, go to Start, Programs, Administrative Tools
(Common), and the click on User Manager.
4. Click on the Policies menu and select User Rights. The User Rights Policy
window will display. In the Right: field, use the drop down list to select
Change the system time. Click Add... The Add Users and Groups window
displays. In the Names: field, select QuantaSmart User. Click Add.... Click
OK. Click OK.
Administrator
in the User: field. Leave the
5. Exit out of the User Manager program.
6. Go to Start, Shutdown, click on Restart the computer?. Click on Yes.
7. When the Windows NT logo displays, press Shift steadily until the Logon
Information window displays. Type
QuantaSmart
in the User: field. Type QS
(this is case sensitive) in the Password: field. Click OK.
Note:You can also access the Date/Time Properties window by
going to Start, Settings, Control Panel, and then double
clicking Date/Time.
Double click on the date and time on the task bar. The Date/Time Properties
window will display. Use this window to change the date and time.
32 PerkinElmer Life and Analytical Sciences
THE SYSTEM COMPUTER
Chapter 3
The System Computer
The system is controlled by an IBM-compatible Pentium® computer. The standard
computer is configured with one floppy disk drive, a CD-drive and one hard drive.
The computer contains a serial RS-232 communications port, a parallel printer
port, and a graphics adapter for the color monitor. All RS-232 communication
occurs through communications port one. Communications port two is used for
proprietary purposes and cannot be used in conjunction with any peripheral
devices.
Attaching To A Network
The system can also be attached to a network. We sell a networking kit to attach
the system to a network. For additional information on the networking kit, contact
your PerkinElmer representative.
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CHAPTER 3
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Chapter 4
The System Software
The QuantaSmart™ program is the Windows XP interface for the TriCarb Liquid
Scintillation Analyzer series of instruments. This innovative tool allows you to take
advantage of all the instrument features via a main software window. The main
window uses standard Windows conventions and allows you to easily access
and control all the system features and capabilities. It also provides you with
graphical representations of existing assay associations in the Protocols Tree and
data analyses in the Output Report and SpectraView windows. Any assay data
that is collected may be reanalyzed, without recounting samples using the
system’s optional Replay feature.
The system utilizes a set of Libraries to store standard and sample information.
For systems equipped with the single/dual label DPM feature, the Nuclide Library
allows you the flexibility of selecting and using the same standards sets in any
number of different assays. It also provides you with a convenient means of
saving and reusing specific nuclide parameters and sample counting regions.
Software Security
Security is built-in to QuantaSmart. The same software cannot be loaded on
multiple systems due to this security feature.
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Main Window
The main software window is comprised of several functional elements which
provides you with a means of accessing all instrument features.
Figure 4-1 Main Window
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THE SYSTEM SOFTWARE
Output Window
The Output Window is a multipurpose window which displays various data items
and assay parameters. It is a copy of the printer report if one is selected. The
output is defined in the Report Output window.
The following is a typical Output Window display:
Figure 4-2 Output Window
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The SpectraView Window
The SpectraView window is part of the main window. It displays a twodimensional, real-time view of the spectrum for the current sample. The spectrum
is updated every six seconds and can be displayed using either linear or
logarithmic axes. It provides you with inf ormation about the status of a sample
count and the region settings used in the counting procedure. A number of
display options are available for the spectrum and are defined in this window.
Figure 4-3 SpectraView Window
The SpectraView window is typically used for the following:
1. Monitoring sample counting.
2. Detecting spectral distortions or compressions resulting from sample quench.
3. Observing the effect of altering the counting region settings.
4. Viewing the spectrum in linear or logarithmic scale.
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The Instrument Status Bar
The Instrument Status Bar contains a series of graphical buttons which allows you
to Stop and Start the instrument and end a current protocol. It also provides you
with information regarding the status of a current protocol and displays
instrument messages.
Figure 4-4 Instrument Status Bar
The buttons in the Instrument Status Bar of the main software window allow you
to start, stop and end a counting procedure.
Stop Button
Click this button to end the current protocol and stop the instrument.
Count Button
Click this button to begin a counting protocol.
End Protocol Button
Click this button to end a counting protocol and continue counting the next
protocol.
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The Protocols Tree
The Protocols Tree displays up to sixty available protocol flag numbers and the
assay names that you have associated to flag numbers. Existing reports are also
shown in this view. During protocol execution, this window uses dif ferent symbols
to provide a visual indication of which protocol is being executed, which protocols
have remaining cycles, and which protocols have been completed.
Figure 4-5 Protocol Tree
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THE SYSTEM SOFTWARE
Symbols Used in the Protocols Tree
When a sample counting cycle begins, the protocol flag associated with this assay
will change from white or yellow to green.
Figure 4-6 Protocol Tree Symbols
The Priostat flag is the only flag in the Protocols Tree that is normally red. It will
turn from red to green when the Priostat protocol begins.
Figure 4-7 The Priostat Flag
A yellow flag in the Protocols Tree indicates that at least one count cycle for this
assay has been completed but that the protocol has remaining count cycles. This
yellow flag will also appear if you interrupt the protocol for a Priostat operation.
Figure 4-8 The Yellow Flag
A checkered flag is displayed when all the count cycles for the assay are
completed.
Figure 4-9 The Checkered Flag
A red prohibitory symbol indicates that a protocol cannot be counted. Typically,
this will result when no assay is associated with a protocol flag number that the
instrument has detected. This symbol could also indicate that an assay file has
been deleted.
Figure 4-10 The Red Prohibitory Symbol
A yellow prohibitory symbol indicates that data analysis cannot be performed.
Typically, this will result when an appropriate standard set is missing from the
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Nuclide Library, or it has been modified in the Library but not recounted after the
changes were made.
Figure 4-11 The Yellow Prohibitory Symbol
The page symbol indicates that a report has been defined for this assay. The
report name appears next to this symbol in the Protocols Tree.
Figure 4-12 The Page Symbol
A white flag indicates an inactive protocol. If no assay name appears adjacent to
this flag, it is available to be associated to an assay.
Figure 4-13 The White Flag
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THE SYSTEM SOFTWARE
The Replay Tree
Replay (optional on 2800; standard on 2900, 3100 and 3170) displays a directory
of folders containing previously collected data which can be reanalyzed using
different data reduction conditions, without recounting the samples.
Figure 4-14 Replay Tree
Note: Clicking twice on the Results folder will bring up the Replay
tree.
The Replay files have a fixed data storage path, but it can be changed in the
Report Output window. The default filename is autoincrementing.
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Menus
At the top of the QuantaSmart main window is the Menu Bar. The Menu Bar
consists of the File, Run, View, Libraries, Tools, IPA, Diagnostics, Window and
Help menus, each of which offers a variety of selections and commands. Make
menu selections by using the mouse pointer to select a menu bar item. Click on
an item to display a list of options within that menu. Note: Some menu items
represent optional features. If your instrument is not equipped with the optional
features, the corresponding menu items will be disabled.
Figure 4-15 Menu Bar
Individual menus can also be displayed by using the Alt-key in conjunction with
the underlined character shown with each menu in the menu bar. Once a menu is
displayed, you may display a menu item by pressing the underlined character key
indicated. For example:
Use the Alt-T keys to display the Tools menu:
Figure 4-16 Tools menu
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File Menu
Display the File menu by selecting File from the Menu Bar.
Figure 4-17 File Menu
New Assay
This menu item allows you to define a new assay.
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Select Assay Type
Before defining the new assay, you must select an assay type in the Select Assay
Type window.
Figure 4-18 Select Assay Type
Open Assay
Select this menu option to open an existing assay.
Associate Assay
This menu item allows you to Associate (link) an assay that you have defined to a
protocol number.
Disassociate Assay
Y ou can disassociate an assa y fr om a pr otocol flag number by selecting the assay
you would like to disassociate in the Protocols Tree of the Main Window. Then
select Disassociate Assay from the File menu. The name of the assay should
disappear from the Protocols Tree. You may also right click on the assay and
select Disassociate Assay from the menu that is displayed.
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Data Paths Window
The Data Paths window allows you to control the location for storing data files
generated by the assay. You may choose this location based on additional
applications that you want to use to process the data or simply to comply with a
data storage strategy in your facility. You can choose to save data to the default
directory by checking the Use Default Output Data Path box.
Figure 4-19 Data Paths Window
Note: This information is stored on a per protocol basis, as
indicated in the Windows title bar of the window.
Print Setup
This menu item allows you to define print parameters.
Print Assays
T o print a list of parameters defined for an assay, select the Print Assays item from
the File menu. Select the assay you would like to print from the Select Assays to
Print window and click Open.
Exit
This menu item allows you to close the QuantaSmart program.
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Run Menu
Display the Run menu by selecting Run from the menu bar.
Figure 4-20 Run Menu.
The Run menu can also be displayed by selecting the Alt-R keys. Each item in the
menu can be displayed by selecting the underlined character indicated.
Stop Counting
This menu item ends the current protocol and stops the instrument. This
command is also available via the red button on the Instrument Status Bar.
Start/Resume Counting
This menu item loads the sample currently at the detector position and begins
counting. This command is also available via the green button on the Instrument
Status Bar.
Next Sample
This menu item unloads any sample in the detector, moves the next sample into
the detector and starts counting.
Next Protocol
This menu item unloads any sample in the detector and aborts the current
protocol. The instrument searches for the next cassette with an active protocol
flag and begins running that protocol.
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THE SYSTEM SOFTWARE
End Protocol
This menu item unloads any sample in the detector and ends the protocol; data
reduction continues until data from the last counted sample is reported. The
system begins counting the next protocol. This command is also available via the
checkered button on the Instrument Status Bar.
Forward
This menu item unloads any sample in the detector and moves the sample
changer in a counterclockwise direction.
Reverse
This menu item unloads any sample in the detector and moves the sample
changer in a clockwise direction.
Group Priostat
This menu item allows you to count a set of high priority samples immediately,
while temporarily interrupting the current protocol.
Stop Group
This menu item terminates counting of the Priostat (priority) samples; and
resumes counting of the interrupted samples.
Sample Priostat (not available on the 2800, optional on the 2900 TriCarb)
This is a special interrupt mode which allows you to: Preview sample count rates,
determine optimal counting conditions for variable and cons tant qu ench samp les,
identify an unknown nuclide in a sample and determine the duration of
luminescence. This menu has the following options.
Decay
This menu item allows you to assess the duration of luminescence in a radioactiv e
sample via a decay histogram.
SPC Decay
This menu item allows you to assess the duration of luminescence in a nonradioactive, luminescent sample via a decay histogram.
Identify Nuclide
This menu item allows you to identify an unknown nuclide in your sample using
the Quench Indicating Parameters SIS and tSIE.
Optimize Regions
This menu item allows you to optimize sample counting regions to provide the
highest figure of merit for Normal count mode.
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Reverse Region
This menu item allows you to re-optimize sample counting region settings for a
sample and determine the equivalent unquenched region settings for variable
quench samples.
Low Level Optimize
This menu item allows you to optimize sample counting regions to provide the
highest figure of merit for Low Level count mode.
Alpha Beta Preview
This menu item allows you to view the sample spectrum in the Alpha/Beta mode
and approximate the activity for a sample containing both an Alpha and a Beta
emitting nuclide.
Normal Preview
The menu item allows you to view the sample spectrum and approximate the
activity for a sample using Normal count mode.
Low Level Preview
This menu item allows you to view the sample spectrum and approximate the
activity for a sample using Low Level count mode.
View Menu
The View menu can also be displayed by selecting the Alt- V keys. Each item in the
menu can be displayed by selecting the underlined character indicated. View has
the following options.
Figure 4-21 View Menu.
Instrument Status Bar
This menu item allows you to view or hide the Instrument Status Bar and its
buttons. The Instrument Status Bar is located underneath the Menu Bar in the
main window.
Status Bar
This menu item allows you to view or hide the Status Bar located at the bottom of
the main window.
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Refresh Trees
This menu item allows you to update the Protocols and Replay trees in the main
window. The Replay tree is automatically updated whenever a new assay is
counted. Refresh Trees is only necessary to clear report entries from the tree
displays.
Libraries Menu
Figure 4-22 Libraries Menu
The Library Concept
Radionuclide information is stored and accessed in the Nuclide Library. The
Nuclide Library consists of the Quench Standards and Sample Nuclides Libraries.
If your instrument is equipped with an Alpha Beta option, an Alpha Beta
Standards and an Alpha Beta Nuclides Library will also be included.
The Quench Standards Library (requires single/dual/color DPM option on the
2800TR) is comprised of quench sets, with each quench set containing individual
quench standards. The data from the quench standards is used to construct
quench curves for calculating DPM (Disintegrations Per Minute) in DPM Assays.
Quench Standards are counted once and the entire spectrum for each quench
standard is stored independent of assay information. This allows you to select and
use the same quench set in any number of assays and construct a quench curve
for each sample at the time the sample is counted.
The Sample Nuclides Library allows you to specify and save nuclide names,
counting region limits and quench sets for sample nuclides. Up to three nuclides
can be defined for each entry to support the counting of multiple nuclides. These
sample nuclide parameters are typically specified as part of the assay definition
process and may be edited as needed.
The Alpha Beta Standards and Alpha Beta Nuclides Libraries (not available on the
2800TR) are used in the same manner as the Quench Standards and Sample
Nuclides Libraries. The information stored in these libra ries is relevant only when
performing Alpha Beta Assays, where both an Alpha-emitting and a Beta-emitting
radionuclide are quantified independently within the same sample vial.
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Sample Nuclides
This menu item allows you to display the Sample Nuclides Library window. The
Sample Nuclides Library is a repository of information regarding sample nuclides.
Figure 4-23 Sample Nuclides window
The Sample Nuclides window allows you to enter information into and retrieve
information from the Sample Nuclides Library.
For assays, use the Sample Nuclides Library to define and save nuclide names and
counting region limits for radionuclides. Y ou can also select a quench set for each
sample nuclide using the Quench Set buttons in this window.
In Replay, use the Sample Nuclide Library to select a radionuclide for the purpose
of reanalyzing sample data.
Note: The fields that are enabled in the Sample Nuclides Library
will be different when accessed from different locations within the
software. The list of nuclides that is displayed is dependent on the
assay type, the nuclide and the number of quench sets associated
with the nuclide. See the Libraries chapter for information on
adding a sample nuclide name.
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Quench Standards
The Quench Standards window allows you to define a new quench set to be
stored or select a quench standards set for DPM assays or data reanalysis in
Replay. The data from these quench sets is used to construct quench curves for
determining sample DPM (Disintegrations Per Minute).
Note: Name, max keV and DPM are the only entries required when
entering a new quench set. The remaining parameters stored in
the library are retrieved from the assay parameters chose in
defining an assay. The number of standards (up to twenty) is
determined automatically during counting.
For additional information, refer to the Libraries chapter.
Figure 4-24 Quench Standards Window
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Alpha Beta Nuclides (not available on the 2800TR)
This menu item allows you to display the Alpha Beta Nuclides Library window. The
Alpha Beta Nuclides Library is a repository of information regarding previously
defined Alpha and Beta emitting nuclides counted in Alpha Beta Assays.
For additional information, refer to the Libraries chapter.
Figure 4-25 Alpha Beta Nuclides Library Window
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Alpha Beta Standards
This menu item allows you to display the Alpha Beta Standards Library window.
The Alpha Beta Standards Library is a repository of information regarding
previously defined standards used in Alpha Beta Assays.
For additional information, refer to the Libraries chapter.
Figure 4-26 Alpha Beta Standards Library Window
Tools Menu
Figure 4-27 Tools Menu.
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CHAPTER 4
Nuclide Decay
Select this menu option to display the Radionuclide Decay window for calculating
the Disintegrations Per Minute (DPM) of radionuclides.
Figure 4-28 Radionuclide Decay Calculator
Options
Select this menu option to display the Options window. The Options window
allows you to select a format for the Spectrum Files cr eated in High Sensitivity and
Low Level Count Modes and activate the feature which allows negative CPM and
DPM reporting.
Figure 4-29 The Options Menu
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Show Protocol Errors
Use this menu item to display certain error messages that are associated with an
assay.
Figure 4-30 Show Protocol Errors
Spectral Mapping
The Spectral Mapping menu item allows you to view a three-dimensional spectral
map for a sample and quench standards. This option is used for single-label DPM
samples.
Figure 4-31 Spectral Mapping
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Spectrum Unfolding
The Spectrum Unfolding menu item allows you to view a three-dimensional
display of the separated, individual spectra f or each nuclide in a Dual-label or Full
Spectrum (FS) DPM sample counting procedure.
Figure 4-32 Spectrum Unfolding.
IPA Menu
Display the IPA™ (Instrument Performance Assessment) menu by selecting IPA
from the menubar.
Figure 4-33 IPA Menu
The IPA menu can also be displayed by selecting the Alt-P keys. Each item in the
menu can be displayed by selecting the underlined character indicated.
IPA Definition
This menu item allows you to define the parameters used to assess instrument
performance.
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IPA Charts &Tables
This menu item allows you to view, edit and print all instrument performance
assessment parameters in chart or tabular form.
Figure 4-34 IPA Charts & Tables.
Diagnostics Menu
Display the Diagnostics menu by selecting Diagnostics from the menubar. The TSE
Diagnostics item in the Diagnostics menu is for the use of a PerkinElmer Service
Engineer to view the system’s diagnostic screens and assess the system’ s
functional status. A password is required.
The Diagnostics menu can also be display ed by selecting the Alt-D k eys. The item
in the menu can be displayed by selecting the underlined character indicated.
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Window Menu
Display the Window menu by selecting Window from the menu bar. This menu
allows you to define a format for the window display on the monitor and restore
the SpectraView window.
Figure 4-35 Window Menu
Cascade
This menu item displays the open windows in the following fashion:
Figure 4-36 Cascaded Windows
Tile
This menu item displays the open windows in the following fashion:
Figure 4-37 Tiled Windows
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SpectraView
SpectraView is a window in the QuantaSmart program which displays a twodimensional, real-time view of the spectrum for the current sample.
Help Menu
Display the Help menu by selecting Help from the menu bar.
Figure 4-38 The Help Menu.
Help Topics
This menu item will launch the On-Line Help documentation with the Table of
Contents initially in view.
About QuantaSmart
This menu item indicates the version of the QuantaSmart software you are using
and the date and time of its creation.
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Spectral Displays
Spectral Mapping
During a single-label DPM sample count, the Spectral Mapping window can display
the sample and quench standards spectra in a three-dimensional view . The X-axis
of the map represents the energy in keV, the Y-axis represents the counts and the
Z-axis represents the quench indicating parameter, tSIE. The spectral map can be
used for the following:
Comparing a sample spectrum to the quench standard spectrum.
Checking for spectral anomalies.
Display a spectral map by selecting the Tools-Spectral Mapping menu option.
Figure 4-39 Spectral Mapping Window
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A sample with a quench level exceeding the limits of the quench standards will
have its position on the map determined by extrapolation. The lower limit of
extrapolation is the tSIE plus 10% of the least quenched standard; the upper limit
of extrapolation is the tSIE minus 10% of the most quenched standard.
Spectrum Unfolding
During a Dual-label DPM or Full Spectrum DPM sample count, the Spectrum
Unfolding window can display three dimensionally, the composite nuclide
spectrum as individual, separate spectra for each nuclide. The X-axes of the
spectral displays represent the energy (in keV) for each counting channel up to
the defined endpoint of the sample spectrum. The Y-axes represents the gross
counts for the current sample. Spectrum Unfolding is often useful for the
following:
Visualizing the relationship of the individual nuclide spectra.
Approximating the ratio of lower energy nuclide to higher energy nuclide.
Display the unfolded spectra by selecting the T ools-Spectrum Unfolding menu
option.
Figure 4-40 Spectrum Unfolding Window.
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The SpectraView Window
The SpectraView window is part of the main window. It displays a twodimensional, real-time view of the spectrum for the current sample. It provides
you with information about the status of a sample count and the region settings
used in the counting procedure. A number of display options are available for the
spectrum and are defined in this window.
Figure 4-41 The SpectraView Window.
The SpectraView window is typically used for the following:
Monitoring sample counting.
Detecting spectral distortions or compressions resulting from sample quench.
Observing the effect of altering the counting region settings.
Viewing the spectrum in linear or logarithmic scale.
The X-axis of the spectral display represents the energy (in keV) for each counting
channel up to the defined endpoint of the sample spectrum. The Y-axis represents
the gross counts for the current sample. The regions settings are graphically
displayed using solid line boxes. This window is updated every six seconds.
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Reports
There are a variety of reports that you can generate using the QuantaSmart
system, either printed or electronic. On the Report Definition tab of the Assay
Definition window, first define the content and name the report(s) that you
need. After creating one or more named reports, go to the Report Output tab to
choose how each named report is to be generated.
Printed Reports
There are a variety of reports that you can print using the QuantaSmart™ system.
On the Report Definition tab of the Assay Definition window, first define a
named report with the information that you need. On the Report Output tab,
select the desired named report and mark the Output to Printer check box. The
printed report(s) you define for an assay will automatically print after the assay is
completed.
To print additional reports, select the report you would like to print from the
Protocols tree in the main window. Click the Print button to print the report when
the Output window is displayed.
To print a list of the parameters you have selected for an assay, select the FilePrint Assays menu option. The Select Assays to Print window is displayed. Select
an assay that you would like to print and click OK.
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Electronic Reports
There are a variety of electronic reports that you can generate using the
QuantaSmart™ system. On the Report Definition tab of the Assay Definition
window, first define a named report with the information that you need. On the
Report Output tab, select the desired named report and indicate what type of
electronic report that want to generate. The electronic output formats include:
Rich Text Format (RTF)
Delimited Text (ASCII)
RS232 (cabled transmission, not a file format)
Other, predefined sets of data can also be selected for output using the Special
Files tab of the Assay Definition interface. This other data includes:
Composite Spectra File
Individual Spectrum Files
IPA™ Data
Protocol Data (PROT.DAT)
Protocol Data (2000CA.DAT)
All of the data files are stored to the location that you define for each protocol.
When you associate an assay with a protocol flag for the first time, you will be
prompted for data path information. The Data Paths window appears so that
you can identify information specific to this protocol (not the assay) for data
storage. You can also view or modify this information for existing protocol
associations by choosing the Data Paths item on the File menu.
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ASSAYS
Chapter 5
Assays
CPM Assays
A CPM assay provides you with information regarding the total quantity of
radioactivity within a sample, in one, two or three predefined counting regions.
The data generated is expressed in CPM (Counts Per Minute). CPM reflects only
the activity that is detected, without regard to counting efficiency or sample
interference, such as quench.
Performing a CPM Assay
The following tasks are required when performing a CPM Assay.
Calibrate the instrument, if necessary.
Create a new assay, choosing CPM as the assay type and define the new
assay parameters, OR open an existing CPM assay and edit or review if
necessary. Save the assay in the Assays folder of the Packard\TriCarb
directory.
Associate (link) the assay parameters with a protocol and attach the
corresponding protocol flag to the first cassette to be counted.
Load the cassette(s) with vials and load the instrument with cassettes.
Start the instrument.
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DPM Assays
A DPM assay allows you to quantitate a nuclide or nuclides within a sample. The
data is expressed as DPM (Disintegrations Per Minute). When DPM are being
calculated, the sample must be checked for quench. If a sample is not corrected
for quench, erroneous DPM results may be reported.
For each sample in a DPM assay, the instrument:
Measures the activity in a sample vial in Counts Per Minute (CPM).
Determines the level of quench via one of the Quench Indicating Parameters
(QIPs).
Interpolates the counting efficiency from a quench curve (plots % Efficiency
vs. QIP).
Calculates DPM, where DPM=CPM/Efficiency.
Depending on the TriCarb model, four different DPM assays are available:
Single, Dual and Triple DPM Assays allow you to count one, two or three nuclides
in a sample using one, two and three defined counting regions.
FS DPM (Full Spectrum DPM) Assays allow you to count two nuclides in a sample
using the full spectrum of the sample (no defined counting regions).
Performing a DPM Assay
The following tasks are required when performing a DPM Assay.
Calibrate the instrument, if necessary.
Define and run a Quench Standards Assay so that counting efficiency and
DPM can be determined for the samples.
Create a new assay choosing DPM Single, Dual or Triple as the assay type
and define the new assay parameters.
Save the assay, OR open an existing DPM assay and edit or review if
necessary.
Select a quench standards set for use with the sample nuclide(s).
Save the assay in the Assays folder of the Packard\TriCarb directory.
Associate (link) the assay parameters with a protocol and attach the
corresponding protocol flag to the first cassette to be counted.
Load the cassette(s) with vials and load the instrument with cassettes.
Start the instrument.
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ASSAYS
Quench Set
The Quench Set field is displayed only in DPM Assays. This field displays the
quench set you selected for the Sample Nuclide in the assay. When using a
quench set, counting efficiency is determined for each sample in the assay. By
selecting the Constant Quench option in this field, counting efficiency will only be
determined for the first sample in the assay. The counting efficiency for this
sample is used to calculate DPM for all the samples in the assay. The Constant
Quench option is advantageous since it results in a reduction in the amount of
time required to count the samples in an assay.
FS DPM Assay
The Full Spectrum DPM method is used for regionless counting of dual label
samples. It uses the Quench Indicating Parameters, Spectral Index of the Sample
(SIS) and transformed Spectral Index of the External standard (tSIE), as well as a
spectral unfolding technique to separate the composite spectrum of the sample.
The composite spectrum is separated into two component spectra, each of which
is the result of a different nuclide. Spectrum unfolding yields the actual CPM for
each nuclide and a quench correlation curve comparing SIS vs. tSIE and Efficiency
vs. tSIE is used to determine the DPM of the unknown samples.
Maximum accuracy with this method is obtained when the concentration of
nuclides in the sample falls in the range of 50:1 to 1:8 (low energy to high energy
nuclide). The quench standards used for FS DPM should be of the same chemistry
and geometry as your unknown samples.
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Performing an FS DPM Assay
The following tasks are required when performing an FS DPM Assay.
Calibrate the instrument, if necessary.
Define and run a Quench Standards Assay so that counting efficiency and
DPM can be determined for the samples.
Create a new assay choosing FS DPM as the assay type and define the new
assay parameters.
Associate (link) the assay parameters with a protocol and attach the
corresponding protocol flag to the first cassette to be counted.
Load the cassette(s) with vials and load the instrument with cassettes.
Start the instrument.
Save the assay.
Or
Open an existing FS DPM assay and edit or review if necessary.
Select a quench standards set for use with the sample nuclide(s).
Save the assay in the Assays folder of the Packard\TriCarb directory.
Associate (link) the assay parameters with a protocol and attach the
corresponding protocol flag to the first cassette to be counted.
Load the cassette(s) with vials and load the instrument with cassettes.
Start the instrument.
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ASSAYS
Direct DPM Assays
The Direct DPM Assay
The Direct DPM Assay is a predefined assay which performs the DPM calculation
based on the Quench Indicating Parameter, SIS (Spectral Index of the Sample).
Since this assay comes pre-installed as part of the system, there is no need to
define individual assay parameters. This assay will calculate accurate DPM values
for single label beta or beta/gamma nuclides, including Tritium. Single-label
samples containing different beta nuclides may be counted in the same cassette.
When using the Direct DPM method, samples are evaluated as follows:
If the SIS value is greater than or equal to 40, then the DPM is reported with
no message on the printout.
If the SIS value is between 20 and 40, then the DPM is reported and the
sample is designated as indeterminate (I) on the printout.
If the SIS value is less than or equal to 20 and the sample is determined to
contain Tritium, the DPM is reported with no message on the printout.
If the SIS value is less than or equal to 20 and the sample is determined not
to contain T ritium, then the DPM is not reported and the sample is designated
as indeterminate on the printout.
Note: When a sample is designated as indeterminate on a printout,
the DPM reported may be valid if the sample is not heavily
quenched. Check the tSIE value of the sample to determine the
level of quench. If the tSIE is greater than 200, the DPM reported
is most likely accurate, within statistical counting error. The
accuracy is independent of cocktail density variation, vial size or
type, sample volume, color and chemical quench. Direct DPM is not
recommended for any background level samples counted for a
short time.
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Performing a Direct DPM Assay
The following tasks are required when performing a Direct DPM Assay.
Calibrate the instrument, if necessary.
Create a new assay, choosing Direct DPM as the assay type.
If you are counting samples using PerkinElmer Ultima Gold™, scintillation
cocktail, you must indicate this in the nuclide library. Doing so will ensure that
the appropriate quench curve will be used to calculate DPM for 3H.
Save the assay in the Assays folder of the Packard\TriCarb directory.
Associate (link) the assay parameters with a protocol and attach the
corresponding protocol flag to the first cassette to be counted.
Load the cassette(s) with vials and load the instrument with cassettes.
Start the instrument.
Alpha Beta Assays (not available on the 2800TR)
An Alpha Beta assay allows you to simultaneously count alpha and beta emitting
nuclides in the same sample vial. In some cases, the emission energies for alpha
and beta nuclides may overlap, making discrimination between the nuclides
difficult. To separate alpha and beta nuclide energies, you must take advantage of
their differing pulse decay times. The TriCarb instruments equipped with the
Alpha Beta feature use the Pulse Decay Analysis (PDA) method of nuclide
discrimination. A time-based Pulse Decay Discriminator (PDD) is used to optimize
the separation of alpha and beta pulses. The optimum PDD value minimizes the
likelihood of alpha events being counted as beta events, and vice versa. To
optimize this setting, you must count a pure alpha and a pure beta standard
source. The optimum discriminator value is where the misclassification of alpha
and beta events is at a minimum. Once the optimum PDD value is established,
you may use this information to count alpha beta assays by referencing the
Standard Set name in the Alpha Beta Nuclide Library. When an alpha beta nuclide
name is chosen in an assay , the referenced Alpha Beta Standard Set name and its
corresponding optimum discrimination is applied in the assay . The data generated
from your sample protocol is expressed in CPM (Counts Per Minute) and reflects
only the activity in the vial without regard to counting efficiency or sample
interference (quench).
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ASSAYS
Performing an Alpha Beta Assay
The following tasks are required when performing an Alpha Beta Assay.
Calibrate the Instrument, if necessary.
Define and Run an Alpha Beta Standards Assay (page 75) to establish the
optimal pulse decay discriminator value.
Define an alpha beta nuclide in the Alpha Beta Nuclide Library. Choose a
Standards Set (second column in the table) from the Alpha Beta Standards
Library to use the discriminator setting from the Standard Set. If you do not
choose a Standard Set, manually enter a discriminator setting directly in the
Alpha Beta Nuclide Library.
Open an existing assay or create a new assay by choosing Alpha Beta assay
type. Select the desired alpha beta radionuclide name on the Count
Conditions tab and define the remaining assay parameters
Save the assay in the Assays folder of the Packard\TriCarb directory.
Associate (link) the assay parameters with a protocol number in the protocol
tree and attach the corresponding protocol clip to the first cassette to be
counted.
Load the cassette(s) with vials and load the instrument with cassettes.
Start the instrument.
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Tips and Techniques for Performing Alpha Beta Assays
1. Several factors influence the discrimination of alpha from beta activity in a
mixed sample. First and foremost is quenching. Alpha beta discrimination
degrades in heavily quenched samples. For best performance, it is
recommended to minimize quench, if possible, by either using smaller sample
volumes or eliminating interferences with sample purification techniques.
2. Alpha Beta standards are counted with a wide open 0-2000 region for beta
activity and 0-1000 region for alpha activity. Optimize counting regions in the
Alpha Beta Nuclide Library for counting unknown samples, if desired.
3. Count the alpha and beta standards used to generate the misclassification
curve in your assay to either confirm the misclassification established when the
Alpha Beta standards were counted or to determine the revised
misclassification that results from using optimized region settings. The new
misclassification must be calculated manually from the observed counts for
both alpha (CPMa) and beta (CPMA or CPMB).
Note:Only one region of interest (CPMa) is defined for alpha
activity. This field is automatically included by default in alpha beta
assays. Beta activity can be reported in 2 regions, CPMA and
CPMB. CPMA is always included by default in the report.
4. High sensitivity (HSCM) or Low Level Count mode (LLCM) can be used in an
alpha beta assay. In this case, DO NOT link an Alpha Beta Standard Name to
an Alpha Beta Nuclide Name in the Alpha Beta Nuclide Library. Since all Alpha
Beta Standards are always
counted with normal count mode by design, linking
an Alpha Beta Standard to the Alpha Beta Nuclide name will automatically
choose normal count mode.
Note:High Sensitivity or Low Level count mode is applied only to
the beta counting and not alpha. Alpha background reduction is
achieved with alpha beta discrimination (Pulse Decay Analysis).
5. To use High Sensitivity or Low Level count mode in an assay, define the Alpha
Beta Nuclide in the library and do not link an Alpha Beta Standard set. Again
the Alpha Beta standards should be counted in the assay to determine the
alpha and beta misclassification.
Note:When counting high energy beta emitters in low level count
mode, it may be advantageous to change the Delay Before Burst
value on the Count Corrections tab (default is 75ns). Adjusting the
value, you can optimize the background reduction for the highest
sensitivity. A value between 150 and 300 is typical. The optimum
must be empirically determined with a representative background
and sample.
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Alpha Beta Standards Assay
An Alpha Beta Standards Assay allows you to count pure beta and pure alpha
standard sources. By counting the beta and alpha standards, you can establish
the optimal Pulse Decay Discriminator value, where misclassification of the beta
and alpha events is at a minimum. Establishing this value allows the instrument to
discriminate between alpha and beta emitting nuclides in your samples in Alpha
Beta Assays.
Performing an Alpha Beta Standards Assay
The following tasks are required to run the Alpha Beta Standards Assay. Two
standards are required, a pure beta emitter and a pure alpha emitter.
Choose the Alpha Beta Standards selection from the Libraries menu.
Click the Add button and enter the name for a new Alpha Beta Standard Set.
Choose Automatic for the Discriminator Type if your pure alpha and pure
beta standards have an activity of at least 50,000 CPM each. If the activity in
either standard is less than 50,000 CPM, choose Manual as the Discriminator
Type by clicking on it.
The remaining fields are for information only. They are either default values
or values computed by the instrument.
Choose File-New and select Alpha Beta Standards as the assay choice from
the drop down menu.
Click the Name button on the Count Conditions Tab to choose from the list of
Alpha Beta Standard names in the Alpha Beta Standards Library. Define the
other available assay parameters as desired.
Note:High Sensitivity or Low Level Count mode are not available
when counting Alpha Beta Standards, but will be available when
counting samples in an Alpha Beta assay
When the assay definition is complete, name and save the assay.
Associate (link) the assay parameters with an available protocol flag in the
protocol tree. Place the numbered protocol clip on a cassette.
Place the pure beta emitter standard in cassette position 1 and the pure
alpha emitter standard in cassette position 2.
Click the green start button at the top of the main window.
After counting is complete, the misclassification (or spillover) curve and the
optimum discriminator value will be stored in the Alpha Beta Standards
Library.
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Quench Standards Assay
A Quench Standards set is composed of a series of vials, each containing the
same amount of nuclide with varying amounts of quenching agent. Using the data
from the quench standards, a quench curve is generated to determine the
counting efficiency for a sample and calculate DPM (Disintegrations Per Minute),
where DPM=CPM/Efficiency. The system stores the spectrum of each standard in
the quench standards set. The quench standards set needs to be counted one
time only, as the quench data is available for use with any protocol.
To accurately assess the level of quench within a sample, the nature and
composition of the quench standards should reflect the matrix and environment of
the samples you would like to count.
Performing a Quench Standards Assay
The following tasks are required when performing a Quench Standards Assay.
Calibrate the instrument, if necessary.
Create a new assay, choosing Quench Standards as the assay type and
define the new assay parameters, OR Open an existing Quench Standards
Assay and edit or review if necessary. Save the assay in the Assays folder of
the Packard\TriCarb directory.
Associate (link) the assay parameters with a protocol and attach the
corresponding protocol flag to the first cassette to be counted.
Load the cassette(s) with vials and load the instrument with cassettes.
Start the instrument.
Note:When using the Low Level count mode, you must not use
quench standards which have been purged free of oxygen with an
inert gas. The oxygen quenching in unpurged standards facilitates
discrimination between background and true beta events.
Unpurged quench standards are available from PerkinElmer Life
and Analytical Sciences.
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ASSAYS
SPC Assay
A Single Photon Counting Assay is an assay which measures the photons emitted
from non-radioactive, luminescent samples. To detect scintillation, modern Liquid
Scintillation Counters use two photomultiplier Tubes (PMTs) to collect virtually all
of the light produced within a sample vial. Each pulse that occurs during the
sample counting time is registered and expressed as Counts Per Minute (CPM).
Events are considered true decay events from the sample if they occur within a
specified coincidence time. If these events do not occur in coincidence, they are
considered random (background) and are not counted.
In an SPC Assay, only one PMT is used. As a result, coincidence cannot be used as
a means of excluding background. Therefore, it is often important to reduce the
instrument background to its lowest possible level. Lowering the high voltage
supplied to the photomultiplier tube typically decreases background and increases
sensitivity in SPC Assays.
Performing an SPC Assay
The following tasks are required when performing an SPC Assay.
Calibrate the instrument, if necessary.
Create a new assay choosing SPC as the assay type and define the new assay
parameters.
Associate (link) the assay parameters with a protocol and attach the
corresponding protocol flag to the first cassette to be counted.
Load the cassette(s) with vials and load the instrument with cassettes.
Start the instrument.
Or
Open an existing SPC assay and edit or review if necessary. Save the assay
in the Assays folder of the Packard\TriCarb directory.
Associate (link) the assay parameters with a protocol and attach the
corresponding protocol flag to the first cassette to be counted.
Load the cassette(s) with vials and load the instrument with cassettes.
Start the instrument.
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Single Photon HV DAC %
The Single Photon HV DAC % field is displayed only in SPC Assays in the Count
Conditions tab in the Regions section. The high voltage supplied to the
instrument’s Photomultiplier Tube is adjustable. Lowering the high voltage
supplied to the PMT typically decreases background counts and increases
sensitivity. The default setting for this device is 70%. The optimum setting will
need to be determined empirically.
The setting is found in the Radionuclide section in the Count Conditions menu.
Defining an Assay
The process of assay definition is central to the use of the instrument software.
Assays are defined using the seven Assay Definition tabs: Assay Parameters,
Count Conditions, Count Corrections, Report Definition, Report Output, Special
Files, and Worklist. Using these seven tabs, for each assay that you define, you
will:
Enter descriptive information about the nature of the assay and the author of
the assay.
Define a sample nuclide in the sample nuclides library if one does not already
exist.
Link the nuclide to the assay.
Link standards to the assay , if necessary.
Specify the appropriate count conditions and count correction factors that the
instrument will use to analyze the samples.
Define the reports you would like the system to generate.
Output desired report options.
Define an optional worklist to designate Positive Identification numbers and
sample names that correspond to sample numbers on a printout, if desired.
The parameters defined within the context of these seven tabs can be saved and
used or edited at your discretion. All of the assay information that you define and
save becomes a functional entity only after it is associated to a protocol number.
These protocols are recognized by the instrument via a protocol flag. This device
contains an encoded, reflective metal which the instrument uses to identify the
protocol number and sample counting parameters that you have defined and
elected to use. The QuantaSmart program enables you to define an unlimited
number of assays and associate them with up to sixty protocols by enabling the
Lock Assay feature.
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ASSAYS
Assay Parameters
The Assay Parameters tab in the Assay Definition window allows you to designate
an author and provide descriptive information for an assay. You may also prohibit
the editing of assay parameters using this window.
Figure 5-1 Assay Definition - Assay Parameters Tab.
Save As
Click this button to save any changes made in the Assay Definition tabs using a
different filename or file location on the disk.
Password
Enter a password if you would like to restrict editing functions for this assay. You
must check the Lock Assay box before you can enter a password in this field.
Author
Enter your name or other identification as the author of the assay. This is an
optional entry.
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Assay Description
Enter descriptive information about the assay. This information is for future
reference.
Date Created
This field represents the date the assay was created.
Date Modified
This field represents the date the assay was last modified.
Lock Assay
Mark this box if you would like to restrict editing functions for this assay. You must
enter a password in the Password field if you would like to lock the assay.
Assay Type
The Assay Type reflects the selection that you made in the Select Assay Type
window.
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ASSAYS
Count Conditions
The Count Conditions tab in the Assay Definition window allows you to define
specific counting parameters for an assay.
Figure 5-2 Assay Definition - Count Conditions Tab.
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CHAPTER 5
Radionuclide: Name
Clicking on the Name button brings up the Sample Nuclide window allowing you
to select a different sample nuclide.
Figure 5-3 Sample Nuclide Window
Count Mode
Select either Normal, High Sensitivity or Low Level from the drop-down list.
Depending on the TriCarb model you are using, the list of available count modes
will be different. Normal mode is the default and works well for most samples.
High sensitivity provides higher sensitivity as a result of the strict criteria used to
exclude background interference. Low Level provides the highest sensitivity
counting for low activity samples due to even stricter criteria for the exclusion of
background interference (with a minimal compromise in counting efficiency).
Quench Indicator
Quench indicators available are: tSIE, tSIE/AEC or SIS. These parameters
measure chemical quenching in your sample.
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ASSAYS
tSIE
(transformed Spectral Index of the External Standard). Using an external Barium133 standard source, this method assigns a numeric value to the quench
associated with a sample. This determination is independent of the quantity of
radioactivity in the sample and its count rate. The lower the tSIE value, the more
the sample is being quenched. A tSIE value of 1000 represents a completely
unquenched sample. Accurate DPM values can be determined for samples with
tSIE values as low as 10. To ensure good count statistics, the external standard is
typically counted to a 0.5% two sigma counting error, where the gross counts
equal 160,000. tSIE is the most accurate of the quench indicator options and is
typically used for low count rate, variable quench, single label samples.
tSIE/AEC
(transformed Spectral Index of External standards coupled to Automatic Efficiency
Correction). tSIE assigns a numeric value to the quench associated with a sample.
As quench varies, the AEC automatically monitors and adjusts the counting region
to exclude unwanted background. This setting is typically used for dual and triple
label experiments with variable quench samples where optimal region settings are
desired.
SIS
(Spectral Index of the Sample) SIS assigns a numeric value to the quench
associated with a sample. The SIS is determined from the spectral shape of the
sample and is based on actual sample counts. The SIS setting is typically used to
monitor the quench level in single label, high count rate samples for CPM assays
or in single label Cherenkov counting.
Note: By using one of the count termination parameters, this count
time may be shortened.
External Standard Terminator
Select a length of time the external standard is counted for calculating the quench
index. Selecting 0.5 2s% instead of an increment of time will allow counting to
occur until gross counts of 160,000 are measured. This provides statistical
accuracy of 0.5% (at 95% confidence) for the tSIE parameter. You may elect to
use the External Standard Terminator only if tSIE or tSIE/AEC are chosen as
quench indicators.
Pre-count Delay
Enter the length of time you would like the samples to sit in the closed detection
chamber prior to counting. This process is “dark adaption”; it will reduce
luminescence originating from the samples. Luminescence can distort the count
statistics of the sample and is particularly problematic with low count rate samples
and long count times.
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Count Time
Enter the maximum length of time that the samples will be counted. For low
activity samples, longer count times provide better count statistics and more
accurate sample results. Changes to this parameter for an active assay will be
implemented immediately.
Assay Count Cycles
Enter the number of times you would like the assay to count. The assay is
recounted after it has moved one complete cycle around the sample changer
deck. Any samples on the sample changer deck will be counted prior to your
samples being recounted.
Repeat Sample Count
Enter the number of times you would like each sample counted while in the
detector. This differs from Assay Count Cycles, where samples are unloaded from
the detector and make a complete cycle around the sample changer deck prior to
recounting.
Calculate Percent Reference
Activate the percent reference calculation by marking this box. The instrument
reports the value of each sample as a percentage of a reference vial. The
reference vial should be the first non-background vial loaded in the cassettes.
Number of Vials per Sample
Enter the number of replicates of each sample being counted. The data output
will report the average value of the replicates.
Lower Limit Regions A, B, and C
These fields represent the lower counting limit for regions A, B, and C, measured
in keV.
Note: This field will only be enabled for Single Label DPM Assays
where tSIE is selected as the Quench Indicating Parameter. The
system uses the tSIE and the sample spectrum endpoint to
determine sample heterogeneity.
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ASSAYS
Background Subtract
Mark this box to subtract background CPM from all samples. The background
value is established in one of three ways and is selected from the following:
1st Vial
The instrument counts the first vial in the cassette for either ten minutes or the
defined protocol count time (whichever is greater) and establishes a CPM value
for each region; these are the background values subtracted from each sample
within each region of the assay.
IPA
The instrument subtracts the background values established during the calibration
and Instrument Performance Assessment (IPA) procedures from the entire
spectrum of the samples. The background spectra are stored during these
procedures and are available for any counting region.
Manual
Enter the CPM values you would like the instrument to subtract from the entire
spectrum of the samples.
Note: In Quench Standards assays, the 1
st
Vial background
subtraction option is not available. Background subtraction is only
applied to the reported data for quench standards and has no
impact on the spectrum for each standard. Any background
subtraction that occurs in DPM assays will apply to the quench
standards used for the purpose of recalculating the quench curve
in the DPM assay.
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Two Sigma Percent Terminator
Mark this box to activate count termination from statistical accuracy. Two options
are available:
Any region
Enter the level of statistical accuracy for each region (as a percent v alue) that you
would like to achieve before counting terminates. Counting terminates when the
sigma value of any one region is reached. Using this feature, counting may
terminate before the specified count time elapses.
All regions
Enter the level of statistical accuracy for each region (as a percent v alue) that you
would like to achieve before counting terminates. Counting terminates when the
sigma value for each region is reached. Using this feature, counting may
terminate before the specified count time elapses.
Changes to this parameter for an active assay will be implemented immediately.
Low CPM Threshold
Mark the check box to activate low CPM count termination. You can enter CPM
values for each counting region to have counting terminate if these values are not
reached. The sample count terminates if any one of the regions does not meet
the specified minimum CPM threshold within the first 30 seconds of counting.
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ASSAYS
Count Corrections
The Count Corrections tab in the Assay Definition window allows you to define
specific count correction parameters for an assay.
Figure 5-4 Assay Definition - Count Corrections Tab.
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Static Controller
Mark this box to activate the instrument’s static-controlling device, which is
designed to reduce static originating on the sample vial. Static discharge can
falsely elevate sample counts by producing non-beta pulses. This device should be
activated in most cases. It’s default value is On. It is especially important in low
humidity conditions, when using plastic vials and when handling vials with latex
gloves. To further reduce the likelihood of generating static:
Maintain a relative humidity level above 40%.
Wipe latex gloves with anti-static wipes before handling vials.
Use the Pre-count delay timer. It delays the counting of each sample allowing
time for static-induced pulses to dissipate.
Note: The Static Controller can cause a decrease in the signal to
noise ratio in SPC Assays. Therefore, it is typically not used in this
assay type. It is especially important in SPC Assays to employ the
above mentioned techniques to minimize the likelihood of
generating static.
Luminescence Correction
Mark this box to activate luminescence correction. The instrument corrects the
data for counts resulting from sample luminescence. This feature is optional on
2800 and 2900 series instruments.
Colored Samples
Mark this box to activate color correction. The instrument will correct the data for
color quench. Typically, this is only required if samples are highly colored. Note:
This field will be enabled only for DPM Assays.
Heterogeneity Monitor
Mark this box to activate a device that monitors and flags heterogeneous samples
(such as samples with phase separation). This feature is not available on the
2800TR. It is optional on the 2900TR.
Note: This field will only be enabled for Single Label DPM Assays
where tSIE is selected as the Quench Indicating Parameter. The
system uses the tSIE and the sample spectrum endpoint to
determine sample heterogeneity.
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ASSAYS
Coincidence Time
Specify the length of time (10-200 nanoseconds) that both PMTs must detect
scintillation events. If scintillation events occur “in coincidence (both events must
occur within the specified coincidence time), these events are considered to be
true decay events from the sample. If these events do not occur in coincidence,
they are considered random (background) and are not counted.
Note: When using solid scintillators, the coincidence time may
need to be extended. The optimal setting must be determined
empirically.
Delay Before Burst
Specify the length of time (75-800 nanoseconds) after the initial pulse (prompt
pulse) that the detector looks for additional pulses (afterpulses). Afterpulses,
which occur after the prompt pulse and delay time interval, indicate that a
scintillation event is due to background. Some scintillators (e.g. PerkinElmer
Ultima Gold) produce slower decaying pulses which may require longer delay
times. When using these scintillators, it is advisable to lengthen the delay time to
retain high counting efficiencies. This is especially important when counting high
energy beta-emitting nuclides. The default setting for this parameter is 75.
Note: This parameter is only accessible with high sensitivity and
low level count modes.
Apply Half-life Correction
Mark this box to activate half-life correction. This feature is typically used when
working with short half-life nuclides. The instrument corrects the sample counts
for half-life decay of the nuclide(s) being counted. The Reference Date and Time
are used to make the decay calculation. The default settings for the Reference
Date and Time correspond to the start of an assay.
Note: In Quench Standards assays, activate this feature only if the
DPM value entered into the Quench Standards Library for the
nuclide has not been corrected for decay . If the DPM value entered
in the library for the standards has been corrected for decay, the
half-life correction feature should not be activated.
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Report Definition
The Report Definition tab allows you to custom design a report.
Figure 5-5 Assay Definition - Report Definition Tab.
Report Name
The Report Name field allows you to assign a descriptive name to a report.
Add Report
Click this button to define a new report for the assay. You can choose to use any
of the named report formats in the list for the different output types identified on
the Report Output tab. You can use a different report format for each of the
output types (Printer, Data File, RS232, Rich Text Format), if desired.
90 PerkinElmer Life and Analytical Sciences
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