Michener TITLE: SECTION:4

Simulated Health Bronchial Washing, Bronchial Brushing

Network and BAL Preparation CLCY-4-15

DIAGNOSTIC LABORATORY STATUS: VERSION: 1 Page 1 of 5

CYTOLOGY MANUAL APPROVED

1.0 Purpose:
 It is the Michener Institute’s policy to provide instruction on the safe and effective
preparation of bronchial washing, brushing and BAL specimens.

2.0 Scope:
 Bronchial washings and brushings are analyzed primarily for the detection of tumour in
suspected cases. Bronchial washing should be transported immediately to the cytology
laboratory.
 Bronchial brushings can either be smeared on a slide and fixed on site or the brush can be
sent in solution to the laboratory.
 BAL specimens are used to study the content and composition of the terminal air spaces.
In addition to the detection of tumour, it is used for the detection of opportunistic
infections in immunocompromised patients. In most instances a portion of the BAL
specimen is also sent to the microbiology department.

3.0 Definitions:
 BAL: bronchioloalveolar lavage

4.0 Responsibility:
 All Diagnostic Cytology faculty and students

5.0 Related Policies/Procedures:

 Laboratory Safety CLCY-2-01
 Centrifugation CLCY-4-02
 Cytocentrifugation CLCY-4-03
 Direct Smear CLCY-4-01
 Summary Table of Non-Gyne Preparation Techniques CLCY-4-07

6.0 Equipment: 6.1 Supplies 6.2 Reagents

 PPE  Disposable test tubes/lids  95% alcohol

 Biological Safety Cabinet
 Centrifuge
 Cytocentrifuge
 Coplin jars
 Microslides or cytospin slides
 Pen/pencil/grease pencil
 Disposable pipettes
 Cytoclips/funnels/lids
 Paper towel

PREPARED BY: APPROVED BY: EFFECTIVE DATE:

Catherine Brown Dr. Peter Bridge April 30, 2015
Revised
 Michener TITLE: SECTION:4
Simulated Health Bronchial Washing, Bronchial Brushing
Network and BAL Preparation CLCY-4-15

DIAGNOSTIC LABORATORY STATUS: VERSION: 1 Page 2 of 5

CYTOLOGY MANUAL APPROVED

7.0
PROCEDURE WORK INSTRUCTIONS RATIONALE
STEPS Bronchial Washing and BAL
7.1 1. Treat all samples as if they were infectious. Every
Getting Started 2. Wear appropriate PPE’s. specimen
must be
3. Turn on Biological Safety Cabinet and allow air to treated as if
stabilize for 5 minutes. they are
4. Take samples from refrigerator/designated area and potentially
place in biohazard hood once they have been infectious.
accessioned. These
specimens
5. Gather all supplies into hood. generally
6. Label two slides with accession number arrive fresh in
7.2 Bronchial 7. Label appropriate centrifuge tube(s) with accession the laboratory.
Washing number of the specimen.
preparation that 8. Decant specimen into the appropriate centrifuge
is very mucoid tube(s).
9. Be sure to balance the amount of specimen in each of
the tubes (same volume and density in each).
10. Make sure lids are secure on each tube.
11. Place centrifuge tube(s) in rotor (bucket) and be sure
to balance the rotor (all spaces need not be filled)
12. Make sure rotor lid is secure.
13. Centrifuge the sample
14. When the centrifuge comes to a complete stop, remove
the centrifuge tube(s) and place in the appropriate
sized tube rack in the Biological Safety Cabinet.
15. Allow the cell sample to settle prior to removing
centrifuge tube lid.
16. Decant supernatant using a disposable pipette or pour Use one
off the supernatant. constant
motion when
17. Bronchial washing sediment may be placed directly on pouring off
a glass slide and spread with a second slide if very supernatant
mucoid.
18. Firmly press both slides together creating a monolayer
of cells and breaking up mucous.
19. Place smears immediately in Coplin jar filled with
fixative (95% alc.) for 30 minutes.

PREPARED BY: APPROVED BY: EFFECTIVE DATE:

Catherine Brown Dr. Peter Bridge April 30, 2015
Revised
 Michener TITLE: SECTION:4
Simulated Health Bronchial Washing, Bronchial Brushing
Network and BAL Preparation CLCY-4-15

DIAGNOSTIC LABORATORY STATUS: VERSION: 1 Page 3 of 5

CYTOLOGY MANUAL APPROVED

20. Label the Coplin jars with the laboratory accession

number and type of sample using a grease pencil.
7.3 BAL
21. BAL specimens are further processed using the BAL
specimens cytocentrifugation technique once the specimen has specimens
been centrifuged and the supernatant has been are generally
decanted. less mucoid
22. Cytocentrifuge the BAL sample. than bronchial
washings
23. Dispose of biohazardous materials into the biohazard
disposal bag.
24. Clean and decontaminate the centrifuge and
cytocentrifuge if using after each use.
25. Remove contaminated gloves and place in biohazard
disposal bag.
26. Wash hands.
27. Specimen is stored in refrigerator for one week before
disposal.

7.0
PROCEDURE WORK INSTRUCTIONS RATIONALE
STEPS Bronchial Brushing
7.1 1. Treat all samples as if they were infectious.
Getting Started 2. Wear appropriate PPE’s.
3. Turn on Biological Safety Cabinet and allow air to
stabilize for 5 minutes.
4. Place samples from refrigerator/designated area and
place in biohazard hood if brush was received in
solution. Go to 7.3 if brush was prepared at bedside
7.2 Bronchial 5. Gather all supplies into hood.
brushing
Preparation
6. Label two slides with accession number
received in 7. Shake the specimen container to remove cells from the
solution brush.
8. The brush may be removed and placed on a labeled
slide in a petri dish.
9. Using a wooden stick forcefully remove material from
the brush and onto the slide.
10. Smear material with the second slide using Direct

PREPARED BY: APPROVED BY: EFFECTIVE DATE:

Catherine Brown Dr. Peter Bridge April 30, 2015
Revised
 Michener TITLE: SECTION:4
Simulated Health Bronchial Washing, Bronchial Brushing
Network and BAL Preparation CLCY-4-15

DIAGNOSTIC LABORATORY STATUS: VERSION: 1 Page 4 of 5

CYTOLOGY MANUAL APPROVED

Smear method.
11. The remaining material in liquid may then be
centrifuged and cytocentrifuged.
12. Place centrifuge tubes in rotor (bucket) and be sure to
balance the rotor (all spaces need not be filled)
13. Make sure rotor lid is secure.
14. Centrifuge the sample
15. When the centrifuge comes to a complete stop, remove
the centrifuge tubes and place in the appropriate sized
tube rack in the biological safety cabinet.
16. Allow the cell sample to settle prior to removing Use one
centrifuge tube lid. constant
motion when
17. Decant supernatant using a disposable pipette or pour pouring off
off the supernatant. supernatant
18. The sediment/cell button is resuspended using a
disposable pipette.
19. Follow the Cytocentrifuge procedure
20. Place slides in Coplin jar with fixative (95% alc.) for
30 minutes.
21. Label the Coplin jars with the laboratory accession
number and type of sample using a grease pencil.
22. Dispose of biohazardous materials into the biohazard
disposal bag.
23. Clean and decontaminate the centrifuge and
cytocentrifuge after each use.
24. Remove contaminated gloves and place in biohazard
7.3 Bronchial disposal bag.
brushing
prepared on site
25. Sometimes the Respirologist will roll the brush
directly onto a slide on site and immediately fix with
cytospray. The slide is labeled with the patient name,
hospital number and location of the brush (ie. LUL,
RML)
26. It is placed into a cardboard jacket for transportation to
the lab with accompanying requisition.
27. The cytotechnologist will check the requisition and
match it to the slide and assign a cytology number.
28. The cytology accession number is written on the slide
and it is ready for staining.

PREPARED BY: APPROVED BY: EFFECTIVE DATE:

Catherine Brown Dr. Peter Bridge April 30, 2015
Revised
 Michener TITLE: SECTION:4
Simulated Health Bronchial Washing, Bronchial Brushing
Network and BAL Preparation CLCY-4-15

DIAGNOSTIC LABORATORY STATUS: VERSION: 1 Page 5 of 5

CYTOLOGY MANUAL APPROVED

29. Wash hands.

30. Remove contaminated gloves and place in biohazard
disposal bag.

8.0 Documentation:
 File requisitions in appropriate lab binder.
 Enter data into Laboratory Daily Log Book.
 Complete all Quality Control forms

9.0 References:
 Bibbo M., Wilbur D., Comprehensive Cytopathology, 3rd ed., Elsevier Inc, 2008.
 The Michener Safety Manual (current version)

PREPARED BY: APPROVED BY: EFFECTIVE DATE:

Catherine Brown Dr. Peter Bridge April 30, 2015
Revised