Olympus 1X71 User Manual

Revised December 2014

UCL Institute of Child Health

User guide

Olympus 1X71

Dr Bertrand Vernay

Light Microscopy Facility Manager

Wellcome building, Office W2.09
Tel Office: 42224

Tel Mobile: [phone]

Email: [e-mail]

Table of contents

page 3 ......... Ownership

page 3 ........ Access Rules

page 3 ......... Olympus Customer Support Contact

page 4 ........ General Specifications

Microscopy Techniques Available
Objectives
Filter Cubes

Camera
Fluorescence Illumination
Pixels to Microns Calibration
Consumables List

page 5 ......... Quick User Guides

Transmitted Light
Epifluorescence
Image Capture

page 6 ......... Halogen Lamp Operation

page 7 .... Kohler Illumination

page 8 ........ Adjusting the Objective Correction Collar

page 9 ........ Prior Lumen200 Metal Halide Lamp Operation

page 10 .. Image Capture with HClmage

page 11 ... Selecting the Right Fluorochrome/Filter Set

page 11 DAPI
page 12 Endow GFP/EGFP Bandpass
page 13 DsRed(TRITC/Cy3)
page 14 Cy5
page 15 Cy7

page 16 ....Prior Lumen200 Spectral Output

page 17 ... Hamamatsu ORCA-R2 Spectral Response

Ownership

Prof. Jane Sowden, Developmental Biology Unit (Purchased in 2011)

Access Rules

 No access without prior training by the Light Microscopy Facility Staff
 Free of hourly charge for Sowden and Ferretti groups, £1 hourly charge for

all other users towards the cost of the consumables is expected

 Prof. Sowden team has priority over other users.
 Users must always record their activity in the Log book
 Problem(s) with the microscope should be reported as soon as they are

noticed

Olympus customer support service: http://www.olympus.co.uk/microscopy

The system is not covered by a maintenance contract
Olympus requires a PO number before sending an engineer

General specifications

Microscopy techniques available



Brightfield



Phase contrast



Epitluorescence

Objectives

 O ly m pu s U P l a n F L N 1 0 x P h 1 NA 0 . 3 W D 1 0 . 0 mm
 Olym p u s LUC P lanFL N 20x P h 1 NA 0 . 45 W D 6.6 - 7.8 w i t h cor r ection collar
 Ol y mpu s L U CP l an F LN 4 0 x P h 2 N A 0 . 6 W D 3 . 0- 4.2 wit h c o rr e cti o n c o lla r

 Z e i s s o bj e ct i ve s c a n a l s o be us e d

Filter cubes (see p11-15)

Position

Filter set name

Exciter

Beamsplitter

Emitter

1

DAPI

350/50x

400LP

ET460/ 5 0m

2

GFP

ET470/ 4 0x

495LP

ET 52 5 / 50m

3

DsRed (TRITC/Cy3)

ET545/30x

T570LPXR

ET620/ 6 0m

4

Cy5

ET620/ 6 0x

T660LPXR

ET700/ 7 5m

5

Cy7

ET710/75x

T760LPXR

ET810/ 9 0m

6

empty (brightfield)

Camera (see p16)

Hamamat su ORCA R2 CCD Came r a with HClma g e Capture So f tware

Fluorescence illumination (see p16)

Prior Lumen 200 Metal Halide Light Source ( 2 0 0 0 h o u r s / b u lb)

Pixels to Microns calibration

5x obje c tive binnin g 1 x 1 1 pixel = 1.88 um
10x obje c tive binning 1x1 1 pixel = 1.03 um
20x obj e c t ive binning 1x1 1 pi x e l = 0.514 um
40x objective binning 1x1 1 pixel = 0.25 6 um

Calibr a t ion with a micrometer unde r tr ansmitted white li gh t

Consumables list

Price correct as of November 2013

 Prior Lum e n 200 bulb LM375 ( £ 550, Prior Scientific Instr u ments Ltd)
 Prior Lumen 20 0 light gu ide LM587 ( £400, Prior Scientif i c Instrum ents Ltd)
 Halogen bulb 12 V/100W (£1 .8, Techn ical Lam p Supplies UK)

Quick user guides

Users must always record their activity in the Log book

Transmitted light

1. H al og en La mp Po we r S u p pl y U ni t TH 4 " O N "

2. L i g ht Pa th se le ct o r on " O cu la r "

3. K o hl er i l lu mi na ti on a dj u s t ed

4. C orr e ct p hase rin g in p osi t ion (10x & 20 x Ph 1 , 40 x Ph2 )

5. F i lt e r c u be o n p o s it io n # 6

Epifluorescence

Warnings:
Do not shut the unit down within 30 minutes of powering up the unit.



After shutting down the unit allow 30 minutes before re-powering up



After shutting down the unit allow 30 minutes before changing the bulb.

Failure to do so is likely to result in damage to the bulb.

1. P rior Lu men 200 mo d ule on

2. P ri or Lu me n 2 00 i n te ns ity k no b > 0%

3. L igh t Pa th s ele cto r le ver on "Oc u lar "

4. C or re c t fi l te r c ub e in p o si ti o n

5. F luo r esc enc e sh utt er o pen

Image Capture

1. S tart Cam e ra c ont r o lle r (pr ess u nti l L ED t u rns gree n)

2. C om put er on ( Lo gi n : Ja n e / P as s wo rd : J a* e)

3. H Cl ma g e so f tw ar e o pe n

4. L ig ht Pa th se le cto r le ve r o n C am er a

5. C orre ct tra n smi tte d li ght / ep i flu ore s ce n ce set - up

6. " L i v e " m o d e

7. Adjust exposure time accordingly. Make use of the Histogram and the Saturatio n

options

8. " A b o r t "

9. " C a p t u r e l "
1 0 . Sa v e a s i n M y D o cu m e n t s > U s e r N a m e _ U n i t > F i l e N a m e . t i f
1 1 . Shut-down: exit HClmage, log out windows session, camera on stand -by (press

until LED turns orange)

Halogen lamp operation:

Turning on the lamp

6

1. Make sure the light intensity control knob

(5)

is in the MIN (minimum intensity)

position on the microscope frame.

2. Make sure the light intensity control knob

(1)

is in the MIN (minimum intensity)

position on the TH4 module.

3. Set the main switch

(2)

to

"I"

(ON) on the TH4 module.

4. On the microscope front, press the transmitted light ON-OFF button

(6)

so that

the button is illuminated.

5. Adjust the brightness with the light intensity control knob

(5).

6. To turn OFF, set the transmitted light ON-OFF button

(6)

to OFF