SKC biosampler 225-9595, biosampler 225-9594, biosampler 225-9595K4, biosampler 225-9597, biosampler 225-9593 Operating Instructions Manual

Operating Instructions

863 Valley View Road, Eighty Four, PA 15330 USA

Tel: [phone] • www.skcinc.com

BioSampler® Bioaerosol Collection Device

Cat. No. 225 Series

Introduction

Description

Inlet

Inlet section

Outlet to pump

Nozzle section*

3 Nozzles (sonic
orifi ces)

Figure 1. SKC BioSampler Components

BioSampler shown with 20-ml vessel (5-ml vessel available)

* Etched nozzle sections and collection vessels are supplied as a matched set. Use the etched identifi er

to match the two components during sampler reassembly for optimal fi t and performance.

Collection
vessel*

Checking BioSampler/Kit Contents

Use the table below to verify that you received all items associated with the Cat.
No. ordered. If you are missing items, contact SKC at 800-752-8472 (U.S. only) or
724-941-9701.

If You Ordered Cat. No. Your Package Should Contain

225-9595

225-9595K4

225-9593

225-9597

225-9594

3-pc glass 20-ml BioSampler (inlet section, nozzle
section, and collection vessel), does not include ground

joint cap; inlet and nozzle sections are a matched set
Four 3-pc glass 20-ml BioSamplers, does not include
ground joint cap; inlets and nozzle sections are each a
matched set

3-pc glass 5-ml BioSampler (inlet section, nozzle section,
and collection vessel), does not include ground joint cap;

inlet and nozzle sections are a matched set

3-pc glass 20-ml BioSampler, two 20-ml collection
vessels (bottoms) with ground joint caps, 1 BioSampler
case with mounting rod, and 1 ViaTrap (120 ml)
collection liquid
3-pc glass 20-ml BioSampler, two 20-ml collection
vessels (bottoms) with ground joint caps, 1 BioSampler
case with mounting rod, 1 ViaTrap (120 ml), 1 Vac-U-Go
Sample Pump (115 V, sonic fl ow), reinforced tubing for
connecting pump to BioSampler, and 1 rotameter

Required Equipment/Media

• BioSampler

• Sample pump, with vacuum gauge or manometer, capable of maintaining

sonic fl ow, such as Vac-U-Go (Cat. No. 228-9605), supplied separately or in
kits; see Checking BioSampler/Kit Contents and Accessories/Replacement Parts

• Collection liquid or coating (see Choose a Collection Liquid/Coating)

• 3/8-inch ID silicone tubing

®

• 1/4-inch ID Tygon

• Liquid trap

• Pipe e

• Aluminum foil or Parafi lm

Tip

For optimal operation and convenience, use BioSampler with the

BioSampler case and mounting rod accessories (available in BioSampler
Mini Kit Cat. No. 225-9597 and supplied in BioSampler Complete Kit
Cat. No. 225-9594).

tubing

2

Operation

Set/Calibrate Pump Flow

The BioSampler nozzle section contains three tangential 0.630-mm nozzles that
act as sonic orifi ces. Flow through the nozzles remains constant at approximately

12.5 L/min when used with a pump, such as the Vac-U-Go, that maintains a pressure
drop of 0.5 atmosphere (15 in Hg) or more across the sampler at normal atmospheric
conditions (sonic fl ow).

1. Prepare the calibration train. (Figure 2)
a. Connect the fl owmeter outlet to the BioSampler inlet.

Inlet

Outlet

20

15

10

LPM AIR

5

2

Rotameter

Valve adjust

Vacuum gauge

15

20

25

InHg
vac

30

ASHCROFT

Air Sampler

Cat. No. 228-9605

SKC Inc. Eighty Four, PA

Sample Pump

(Vac-U-Go shown)

Inlet

10

5

®

Tubing

Figure 2. Calibration Train

Inlet

Tubing

BioSampler

If using the rotameter supplied in Cat. No. 225-9594, use a length of 3/8-inch ID
silicone tubing to connect the rotameter outlet to the BioSampler inlet.

b. Connect the BioSampler outlet to the sample pump inlet.

2. Use the valve adjust on the pump to achieve sonic fl ow (vacuum gauge or
manometer indicates between 15 and 30 inches Hg vacuum).

3. Observe the fl ow rate indicated on the fl owmeter and record as Q

L/min (to

s

be used to calculate sample volume).

Calibration Note

If pump fl ow rate changes by ± 5% between the baseline calibration (fi rst calibration
when sampler is new) and subsequent calibrations, it may indicate a broken or clogged
nozzle. Replace the nozzle section of the BioSampler and re-calibrate. The new calibrated
fl ow rate (with replacement nozzle section) will likely diff er slightly from previous
calibrations (each BioSampler has a unique fl ow rate that is approximately 12.5 L/min).
Use this new fl ow rate as the new baseline fl ow rate for this sampler.

3

Prepare the BioSampler Collection Vessel

Before sampling, sterilize the BioSampler, choose a collection liquid or coating, and
fi ll or coat the collection vessel as follows.

Sterilize the BioSampler

Sterilize the inlet, nozzle section, collection vessel, and ground joint cap in
an autoclave or oven heated within a range of 320 to 356 F (160 to 180 C) for 180
to 240 minutes before each use. This form of sterilization will kill bacteria, fungi,
and DNA.

To prevent fritted joints from sticking together during heating, disassemble
sampler components (three pieces) before placing them in autoclave or oven.

During reassembly, match the etched identifi er on nozzle section with the
identifi er on collection vessels for optimal fi t and performance.

Choose a Collection Liquid/Coating

BioSampler can be used for short-term and long-term sampling. See the following
table for recommended collection liquids and coatings based on the desired sample
time. Contact the laboratory for specifi c requirements, including sterilization.

Desired Sampling Time Recommended

Liquid/Coating

Shorter than 30 minutes
(with liquid)

Shorter than 30 minutes
(with coating)

Longer than 30 minutes Non-evaporating liquid

Sterile distilled water,
physiological saline,
phosphate buffered saline
(PBS), nutrient broth, or
peptone water

Petroleum jelly/hexane
solution coating

such as ViaTrap
or glycerol

Note: ViaTrap® mineral oil
requires sterilization before
use. See Note in Fill or
Coat the Collection Vessel

mineral oil

Notes/Cautions

Do not use water or water­based liquids for sampling
periods longer than 30
minutes or for collecting
hydrophobic particles such
as fungal spores. Contact
laboratory for specifi c
requirements.
Do not use this method for
sampling periods longer
than 30 minutes.
 Collection effi ciency

remains virtually
constant during 8-hour
sampling period.

 Glycerol is not

recommended for
viable microorganism
collection; it can cause
microorganisms to die
from osmotic pressure.

 ViaTrap may not

be suitable for PCR
analysis.

4

Fill or Coat the Collection Vessel

1. Fill the collection vessel with collection liquid. You will need:

• Sterilized collection vessel(s)

• Pipe e

• Sterile collection liquid (can include, but is not limited to, sterile distilled
water, physiological saline, PBS, nutrient broth, or peptone water)

• Aluminum foil or Parafi lm

If using ViaTrap mineral oil as the collection liquid, sterilize it by fi ltering it through a

0.4-μm pore size sterile polycarbonate membrane fi lter. Do NOT autoclave ViaTrap;
heating the oil will make it cloudy.

a. In a sterile environment, aseptically pipet the appropriate amount of sterilized

collection liquid into the sterilized collection vessel.

If using a viscous liquid, such as ViaTrap or glycerol, pipet less than 20 ml of
viscous liquid into the 20-ml collection vessel (< 4 ml inside a 5-ml collection
vessel) to prevent overfl ow of liquid outside the sampler.

b. In the same sterile environment, assemble the clean, sterilized inlet, nozzle

section, and fi lled collection vessel.

c. Temporarily seal sampler inlet and outlet with aluminum foil or Parafi lm to

maintain sterility.

2. Coat the collection vessel. You will need:

• Sterilized collection vessel(s)

• Pipe e

• Sterile solution for coating (can include, but is not limited to, petroleum
jelly/hexane solution)

• Aluminum foil or Parafi lm

a. In a sterile environment, prepare coating solution. Note: SKC recommends

using a petroleum jelly/hexane solution (approximately 0.1 gram petroleum jelly
per milliliter of hexane).

b. Aseptically pipet a few drops of solution into the collection vessel and rotate

it so that its entire inner surface is we ed. Ensure the coating covers the
region where the air jets impact the inner surface of the collection vessel. The
hexane will evaporate within fi ve minutes, leaving a uniform sticky layer on
the inner surface of the collection vessel.

If using the petroleum jelly/hexane solution, the hexane will evaporate within fi ve
minutes, leaving a uniform sticky layer on the inner surface of the collection vessel.
Alternative coatings can be used.

c. In the same sterile environment, assemble the clean, sterilized inlet, nozzle

section, and coated collection vessel.

d. Temporarily seal sampler inlet and outlet with aluminum foil or Parafi lm to

maintain sterility.

5

Set Up/Sample

• If sampling using a collection liquid, especially a viscous liquid such as ViaTrap
or glycerol, SKC recommends using Liquid Trap Cat. No. 225-22 to protect the
sample pump.

• If using a collection liquid, see Proper Operation During Sampling.

• If sampling biologically inert particles, aseptic procedures are not necessary.

• If using a water-based collection liquid, do not sample at temperatures below 41 F
(5 C) to avoid crystallization of the collection liquid.

1. Warm up the vacuum pump for at least one minute before connecting it to the
sampling train. See sample pump operating instructions.

2. When ready to sample, remove foil or Parafi lm from the BioSampler inlet and
outlet.

3. Prepare the sampling train.

a. If using a collection liquid, it is important to use a liquid trap

(Figure 3) to protect the sample pump. Connect the BioSampler outlet
(nozzle section) to the inlet of a liquid trap. Connect the outlet of the
liquid trap to the sample pump inlet.

b. If using a coated collection vessel (Figure 4), connect the BioSampler

outlet (nozzle section) to the sample pump inlet.

Inlet

Outlet

BioSampler

(fi lled with liquid)

Inlet

Tubing

Outlet

Tubing

Liquid Trap

Figure 3. Sampling Train with Liquid Trap

6

Vacuum gauge

20

25

Valve

Sample Pump

(Vac-U-Go shown)

15

10

InHg
vac

30

®

ASHCROFT

Air Sampler

Cat. No. 228-9605

SKC Inc. Eighty Four, PA

Inlet

5

Inlet

Vacuum gauge

Inlet

Outlet

Tubing

15

20

25

InHg
vac

30

ASHCROFT

SKC Inc. Eighty Four, PA

10

5

®

Air Sampler

Cat. No. 228-9605

BioSampler

(coated)

Valve

Sample Pump

(Vac-U-Go shown)

Figure 4. Sampling Train with Coated Collection Vessel

4. Sample for the sampling period appropriate for the collection liquid or
coating used. See Choose a Collection Liquid/Coating. Also see Proper Operation

During Sampling.

5. After sampling is complete, record the sampling time.

6. Calculate volume of air passed through the BioSampler using the following

formula:

V [Liters] = Q

[fl ow rate in L/min] x t [sampling period in minutes]

s

7

Proper Operation During Sampling

If sampling using collection liquid, observe the
sampler frequently during sampling (especially
when using viscous collection liquid) for the
following.

When operating properly, the collection liquid swirls up
the inner wall of the collection vessel, we ing the inner
wall where the nozzles output. Visually monitor swirling
liquid to determine that:

• There is enough collection liquid to wet the

inner wall where nozzles output. If this is
not occurring, pause sampling and add more
sterilized collection liquid before resuming.

Underfi lled

Operation with

properly fi lled

collection vessel

• There is not excessive bubbling, which indicates

too much collection liquid in the collection
vessel. If excessive bubbling occurs, pause
sampling and aseptically pipet some of the
collection liquid from the vessel before
resuming.

Overfi lled

8

Prepare Samples for Transport/Shipment and Analysis

1. In a sterile environment, disassemble the BioSampler.

2. Transfer sample as follows:
a. If collection liquid was used, accurately measure the final volume of

collection liquid and aseptically transfer it into a sterilized container for
analysis.

b. If a coated collection vessel was used, wipe the inner surface of the vessel

with a small piece of cloth or paper. Aseptically transfer cloth or paper
sample into a sterilized container for transport to a laboratory for analysis.

If sealing a collection vessel with a ground joint cap, apply silicone grease to
the ground glass surface on cap to prevent sticking.

3. Seal individual sample containers to prevent contamination during transport.

4. Properly label all containers with sample identifi cation.

5. Transport or ship samples for delivery to a laboratory as soon as possible after
collection, preferably within 24 hours. Many analyses have very short hold
times.

Analyses Information

Samples collected using the BioSampler can be analyzed by the methods described
below.

Analysis Method Used for

Growth culture Quantifi cation and characterization of airborne

bacteria and fungi to the species level

Microscopic Enumeration and limited identifi cation of total

airborne bacteria and fungi

Biochemical assay Measurement of biological compounds such as

endotoxins

Immunoassay Quantification of airborne allergens. Other

methods include fluorescence, enzyme, and
radioimmunoassay.

Polymerase Chain Reaction (PCR)

Note: If using this analysis method, ViaTrap
mineral oil may not be a suitable collection
liquid. Contact the laboratory for information on
the appropriate collection liquid.

Screening of samples for a specifi c genus or
species based on in-vitro replication of selected
nucleic acid sequences

Maintenance

Store components unassembled (in three pieces) to prevent future diffi culty in
separating pieces, and store them in a clean, dry area away from temperature
extremes.

9

References

Macher, J.M., et al., “Sampling Airborne Microorganisms and Aeroallergens,” Air Sampling Instruments for
Evaluation of Atmospheric Contaminants, 8th ed., ACGIH, 1995, pp. 589-617

Macher, J.M., “Evaluation of Bioaerosol Sampler Performance,” Applied Occupational Environmental Hygiene,
12, 1997, pp. 730-736

Bu ner, M.P., et al., “Sampling and Analysis of Airborne Microorganisms,” Manual of Environmental Microbiology,
ASM Press, Washington, DC, 1997, pp. 629-640

Lin, X., et al., “Eff ect of Sampling Time on the Collection Effi ciency of All-Glass Impingers,” Am. Ind. Hyg. Assoc.
Journal, 58, 1997, pp. 480-488

Willeke, K., et al., “Improved Aerosol Collection by Combined Impaction and Centrifugal Motion,” Aerosol Science
and Technology, 29 (5), 1998, pp. 439-456

Lin, X., et al., “Long Term Sampling of Airborne Bacteria and Fungi into a Non-Evaporating Liquid,” Atmospheric
Environment, 1999, pp. 4291-4298

Lin, X., et al., “Survival of Airborne Microorganisms during Swirling Aerosol Collection,” Aerosol Science and
Technology, 32 (3), 2000, pp. 184-196

Willeke, K., and Macher, J. M., “Air Sampling,” Bioaerosols: Assessment and Control, 1999, pp. 11-1 to 11-25

Reponen, T., et al., Biological Particle Sampling. Aerosol Measurement, Principles, Techniques, and Applications,
John Wiley & Johns, Inc, 2001, pp. 751-777

Rule, A. M., et al., “Application of Flow Cytometry for the Assessment of Preservation and Recovery Effi ciency of
Bioaerosol Samplers Spiked with Pantoea agglomerans,” Environmental Science and Technology, 41 (7), 2007, pp.
2467-2472

Nevalainen, A., et al., Bioaerosol Sampling: Aerosol Measurement Principles, Techniques, and Applications, Van
Nostrand Reinhold, New York, 1993, pp. 471-492

Accessories/Replacement Parts

Description Cat. No.

BioSampler Collection Vessel (bottom) and ground joint cap,
for transporting samples

ViaTrap Collection Media,

†

special mineral oil for bioaerosol

sampling

Sonic Flow Pump includes protective housing with vacuum gauge
and valve, supplied without orifi ces or rotameter, AC operation only

Glass Trap, prevents collection liquid from entering the pump,
can be used with or without sorbent 225-22

Trap Sorbent, 200 grams, silica gel/activated charcoal sorbent
mix to remove vapors 225-22-02

Rotameter, 2 to 20 L/min, 3/16-in ID x 5/16-in OD tubing with 1/8
to 3/16-inch adapter installed 320-4A20L

† ViaTrap mineral oil may not be suitable for PCR analysis.

10

20 ml
5 ml

120 ml
500 ml
950 ml

225-9596
225-9596A

225-9598A
225-9598
225-9599

115 V 228-9605

SKC Limited Warranty and Return Policy

SKC products are subject to the SKC Limited Warranty and Return Policy, which
provides SKC’s sole liability and the buyer’s exclusive remedy. To view the complete
SKC Limited Warranty and Return Policy, go to h p://www.skcinc.com/warranty.asp.

Appendix

Performance Profi le

Flow Rate Approximately 12.5 L/min

Medium Water or more viscous liquids such as ViaTrap mineral

oil† or coating. Contact the laboratory for specific
medium.

Method of Operation Air passes through three 0.630-mm tangential sonic

nozzles that output into a swirling fl ow of collection
liquid, thereby entraining bioaerosols in the liquid for
analysis.

Collection Effi ciency Nearly 100% over a wide range of particle sizes;

decreases to approximately 90% at 0.5 μm

Pump High-volume vacuum pump capable of achieving sonic

fl ow; SKC recommends the Vac-U-Go Sample Pump
(115 V)

† May not be suitable for PCR analysis. Contact the laboratory for information on the appropriate

collection liquid.

11

www.skcinc.com

12

Form 37084 Rev 1707