Acid-Fast Direct Smear Microscopy Training Package Section 2

CDC en Español

Acid-Fast Direct Smear Microscopy Training

	Home

Trainers Guide

	Section 1: Overview

	Section 2: Preparation and Checklists

	Section 3: Training Basics

	Section 4: Trainer's Notes and Presentation Slides

	Section 5: Evaluation Tools

Participant Manual

	Participant Home

Additional Resources

	Reference List

	International Lab

Acid-Fast Direct Smear Microscopy Training Package

Section 2: Preparation and Checklists

Introduction

Thorough preparation is essential to the success of your workshop. This section describes the activities that you must carry out in advance of the workshop to assure that the workshop achieves expected outcomes. Information on the basics of training (Section 3) should also be reviewed in advance so that you, as trainer, are adequately prepared to facilitate the workshop.

Participants (Target Audience)

This training package is designed for laboratory workers who perform direct smear microscopy. It is important to have an understanding of your participant's baseline knowledge and fears so that these can be adequately addressed during the workshop. It may be necessary to adjust the time allotted for a particular module of instruction or exercise based on identified need.

Suggested number of workshop participants and trainers

For optimum learning experience and management of the workshop, it is recommended that the number of participants should not exceed 4-6 participants per facilitator. The overall number of participants depends upon the laboratory facility for the practical sessions and the participant's experience in performing AFB microscopy.

The number should be small enough for all participants to be fully engaged, yet large enough for a variety of experiences and viewpoints to be represented. It is equally important to have one trainer for every 4-6 participants to allow participants ready access to trainers during demonstrations and practical exercise.

Participant Notification Letter

Notify participants well in advance of the date, location, and time of workshop. Be sure to include other pertinent information such as payment of per diems, etc.

Breaks and Lunch

Before the start of the workshop, arrange for morning and afternoon tea/coffee breaks, and plans for having lunch on or off-site.

Facilities and Equipment:

For optimal learning experience and ease of managing logistics throughout the workshop, two rooms should be made available:

Classroom for lectures, discussions, and classroom exercises
Laboratory for hands-on practical exercises

Classroom

The workshop can be held in any well-lit, distraction-free classroom with 1) tables and chairs, and 2) conveniently located outlets for computer and projection monitor. To facilitate discussion and interaction among participants, tables should be arranged in a semi-circle, or classroom style giving all participants an unobstructed view of the projection monitor. Avoid overcrowding. Bottled water and glasses should be made available on each table.

Classroom equipment - The classroom should have:

2 flip charts with easel
markers (different colors for flipcharts)
DVD player with TV or monitor
laptop computer
projection monitor (LCD) compatible with computer
extension cord
wastebasket
laser pointer

Classroom supplies:

marking pens
masking tape for posting flip chart
note pads
pens and pencils
calculators

Laboratory

A well ventilated, lit (artificial or natural) laboratory, with running water is required for AFB smear training laboratory. Hand washing facility with soap and water are essential. A work area of 4 square meters per person should be adequate for performing exercises and recording. Additionally, a clean bench top of 75 cm wide and 150 cm long per person is required to perform exercises. Laboratory must have staining sink or rack. Power supply or natural light area for microscopes is required. Supplies must be labeled and stored in shelves or in other storage area in the laboratory. Demonstration of exercises should be conducted in the center of laboratory or in an area where all participants can have direct and uninterrupted view of the activity. Under no circumstances should laboratory look crowded.

Materials and Supplies

Materials and supplies for demonstration and practical exercises:

Make arrangements well in advance of the workshop to procure or secure the necessary materials, supplies, and reagents. Do not forget to arrange for transport of these items to workshop site. The following is a guide to use for assuring you have an adequate number of supplies on hand during the training workshop. The reagents should be those approved for use in the National Tuberculosis Program. Any unused supplies should be held for future workshops.

	Materials and Supplies	# required per participant
Safety Supplies
	Aprons or laboratory coats
	Paper towels
	Soap or hand sanitizer for hand washing
	Bench disinfectant
	Spray bottle for disinfectant
	Gloves
	Container for broken glass
	Biohazard waste containers
	Waste containers
	Biohazard stickers or labels
	Safety glasses
Laboratory Supplies and Equipment
	Spray bottle and scented oil for Module 1 exercise
	Measuring cylinder (100 ml)
	Measuring cylinder (500 or 1000 ml)
	Weighing balance ( 0.1g sensitivity)
	Flasks of 2 liter capacity
	Weighing paper
	Labels for reagent bottles
	Diamond pencils
	Staining rack
	Rack
	Microscope with oil immersion objective
	Immersion oil
	Lens paper or tissue paper
	Spirit Lamp for heating
	Permanent marker
	Containers for newly prepared stain
Specimen Collection
	Sputum cups
	Request form
	Laboratory register
	Marker for sputum cups
Reagent Preparation
	Weighing balance (0.1 g)
	Weigh boats or paper
	Basic fuchsin powder
	Phenol crystals
	Concentrated sulfuric acid of 95% purity
	Denatured alcohol of 95% purity
	Methylene blue powder
	Distilled water
	Spirit Lamp
	Burning alcohol
	Filter funnel
	Cotton and gauze
	Slide panel
Preparation of AFB Smear
	Clean glass slides (frosted end preferred)	10/participant
	Applicator sticks (e.g., bamboo)
	Wire loops and sand alcohol flask
	Ruler marked with centimeters
	Newsprint
	Forceps
Miscellaneous Supplies
	Spray bottle for Module 1 exercise
	Perfume or lavender oil
	Staining racks

Smear and Specimen Requirements

The success of your workshop will depend in large part to the quality of specimens and smears used during the practical laboratory sessions. The practical laboratory session for smear preparation needs to utilize sputum specimens which are similar to those which would be submitted for routine AFB microscopy. However, as this is a training exercise and proficiency of the participants may not have been determined it is important to conduct this practical in a safe manner. Each of the participants should have access to at least three sputum specimens of different consistency to allow the participants develop expertise in preparing smears from all sputum types. Participants will gain maximum benefit from this exercise if they are provided with a purulent, a mucoid and a watery specimen. Specimens should be collected 2 to 3 days prior to the practical to ensure adequate numbers of sputum samples are available. Each pair of participant should be provided with three sputum specimens of different consistency. Options for using sputum specimens are outlined.

Option #	Sputum Type	Description
1	Previously screened and found not to contain AFB.	As the "key" learning objective for this training is the ability to prepare sputum smears of uniform thickness and size. It is not necessary to use sputum specimens that are known to contain AFB. The practical session can be conducted more safely if the participants are provided with sputum specimens that have been previously screened and found not to contain AFB.
2	Sputum specimens pre-treated with formalin	Sputum specimens can be pre-treated with 4 drops of formalin per ml of specimen to kill any AFB that may be present in the specimens. This will allow AFB positive sputum specimens to be smeared safely particularly if the participants are inexperienced.
3	Use known positive sputum specimens	Known positive specimens can be used in the practical sessions if the participants have been previously trained in AFB microscopy and a well ventilated laboratory area is available.

Laboratory Practical Sessions # 1 and # 2: Ziehl-Neelsen Reagents Preparation and Quality Control of Ziehl-Neelsen Staining Reagents Each participant is given 2 unstained smears for quality control of newly prepared stains. QC1 smear should be negative, QC2 smears should be 1+ for AFB
Unstained Smear		# of Demo Participants	+ Extra	Total Number of Smears
QC 1		12	8	20
QC 2		12	8	20
Laboratory Practical Session # 3: Smear Preparation Each participant prepares 10 smears using previously collected sputum specimens. Note: Specimens can be shared amongst participants thereby reducing the total number of specimens to one-half.
Sputum Specimen	QTY	# of Demo Participants	+ Extra	Total Number of Specimens
Purulent	1	12	2	14
Mucoid	1	12	2	14
Watery	1	12	2	14
Laboratory Practical Session # 4: Smear Staining Each participant will stain smears from Laboratory Practical Session # 3 and a set of five smears from the panel provided
Unstained Smear #	Grade	# of Demo Participants	+ Extra	Total Number of Smears
P-1	No AFB	12	8	20
P-2	1+	12	8	20
P-3	2+	12	8	20
P-4	3+	12	8	20
P-5	<9 AFB	12	8	20

Laboratory Practical Session # 5: Viewing of Prepared and Stained Smears from Laboratory Practical Session # 4

Participants' Prepared and Stained smears from Practical Session # 4 will be used during the Practical Session # 5. Participants are to review each smear in terms of

Thickness of smear
Size of smear
Uniformity of staining
Appearance and characteristics of AFB

Results are discussed after participant interpretation.

Laboratory Practical Session # 6: Reading and reporting of Stained Panel Slides from Laboratory Practical Session # 4

Panel Smears Stained in Practical Session # 4 will be used during the Practical Session #6. Participants are to review each smear in terms of

Identify AFB
Counting of AFB
Recording of AFB
Reporting of AFB

Results are discussed after participant interpretation.

Final Practical Examination A total of 10 blinded stained smears are required for each participant.
Participants are asked to evaluate each smear in terms of Reading and interpretation of smear Smear quality For the Final Practical Examination, prepare a panel containing 10 stained smears. For ease of preparation, 4-5 sputum specimens of different grades are required. Slides labeled F-6 to F-10 are a repeat of the first five slides.
Example of Final Practical Examination Each participant reads and interprets using 10 stained smears.
Unstained Smear #	Grade	# participants	Extra
F-1	No AFB	12	8
F-2	1+	12	8
F-3	2+ or 3+	12	8
F-4	< 9 AFB	12	8
F-5	< 9 AFB	12	8
F-6	No AFB	12	8
F-7	1+	12	8
F-8	2+ or 3+	12	8
F-9	< 9 AFB	12	8
F-10	< 9 AFB	12	8

Summary of Smears Required for Final Practical Examination
Grade of Smear	# Slides
No AFB Seem	40
1-9/AFB/100 fields (Scanty)	80
1+	40
2+ or 3+	40

Validation of Panels

All panel samples used during the hands-on sessions MUST be validated prior to the workshop to assure that expected results are obtained on each smear.

Please refer to External Quality Assessment for AFB Microscopy for details on preparing panel of slides of known contents.

Print Materials Needed During Training

As a workshop facilitator or trainer, it is your responsibility to make sure the appropriate materials are available at the start of the workshop for each participant

Pre-course and post-course tests
Forms to record results of practical examinations
Daily evaluation questionnaire
Post workshop evaluation
Final examination form
Participant manual
Name tents
Name badges

Training Certificates
At the end of the workshop, each participant who has met the criteria for successful completion will receive a Certificate of Training. Prior to the workshop, you will need to

Identify appropriate individuals to who will sign the certificate
Verify spelling of names
Print certificates

Training schedule
The theoretical and practical sessions of this workshop are designed to be delivered in 4-5 days.
A template is provided for finalizing your training schedule. Before conducting the workshop, finalize your schedule for the week by completing as follows:

Write in planned times for breaks and lunch.
Add any additional presentations or activities not included in the template.
Fill in planned times for the activities in the Workshop Timing column. For example, if the workshop is scheduled to begin at 8:30 a.m., write 8:30-9:30 a.m. for the Welcoming remarks and introductions.

Duration of Workshop
The duration of any given workshop may depend on several factors:

Availability of facility
Refresher training
Focused training on specific modules
Responsibilities of target audience
Availability of trainers

Click Here for Training Schedule Template

Are You Ready? Checklist

This checklist summarizes all the preparatory activities that must be completed before conducting an Acid Fast Direct Smear Microscopy workshop. If you are not responsible for carrying out a workshop activity, make sure someone else has been assigned responsibility.

ACTIVITY CHECKLIST: PREPARATION FOR ACID FAST DIRECT SMEAR MICROSCOPY WORKSHOP
BEFORE the WORKSHOP
Check When Complete	Activity: Six to Eight Weeks before Workshop
	Adapt course content based on results of needs assessment.
	Verify or develop training budget.
	Identify target audience.
	Agree on maximum number of participants.
	Finalize names of trainers.
	Finalize names of guest speakers for opening and closing ceremonies.
	Send letter of invitation to co-trainers.
	Send letter of invitation to speakers for opening and closing ceremonies.
	Identify who will be responsible for each task : Develop a list of tasks necessary to present course, and negotiate with co-trainers to decide who will complete each task.
	Meet with co-trainers to coordinate roles and responsibilities (announce date, site and agenda in advance of co-trainer meeting).
	Develop and/or review the participant manual and other training material.
	Determine training supplies and materials needed.
	Develop a detailed agenda setting time-frame for course and speakers.
	Set DATE and LOCATION for workshop. Verify that the date does not coincide with major events or religious holidays. Ensure that invited speakers agree on date and place.
Check When Complete	Activity: Four Weeks before Workshop
	Develop workshop announcement and registration materials.
	Mail workshop announcement and registration materials to target audience. Announcement should include: "who should attend", course title, course content/agenda, course objectives, date and location, instructions for registering, lodging/travel information (if necessary) and name of contact person.
	Confirm arrangements for travel and lodging for co-trainers and guest speakers, if necessary.
	Agree on arrangements such as per diem for co-trainers and guest speakers.
	Identify site(s) and contacts for site visit (s), if included in agenda.
	Send letter of confirmation to co-trainers.
	Send letter of confirmation to guest speakers for opening and closing ceremony.
	Send letter of confirmation to hosts of field visit site(s).
	Develop course flyer and registration materials.
	Set agenda for Opening Ceremony and Course Introduction.
	Set agenda for Closing Session.
	Order or assemble training supplies including pencils, pens, reagents, test kits and equipment.
	Provide deadlines for co-trainers for submitting audiovisual materials, audiovisual equipment needs and handouts for printing.
Check When Complete	Activity: Three Weeks before Workshop
	Reserve audiovisual equipment (check working condition, extra light bulbs, and electrical outlet requirements).
	Obtain flip charts, pointers, felt tip markers and other training materials as needed.
	Confirm training venue location. Check venue for adequate light, space, seating arrangement, temperature control, handicap access, etc. If planning a wet workshop, check facility for appropriate space for laboratory supplies and equipment.
	Develop or adapt Pre and Post-test forms.
	Develop or adapt Final examination form.
	Develop or adapt Workshop Evaluation form.
Check When Complete	Activity: Two Weeks before Workshop
	Print and assemble participant manual. Print handouts, Pre-and Post-test forms, Evaluation forms, agenda, etc.
	Check on progress of participant registration.
	Assemble audiovisual materials (power point files, videos, overheads).
Check When Complete	Activity: One Week before Workshop
	Make name tags for participants, co-trainers and speakers.
	Develop sign in sheet for participants.
	Develop course completion form or course certificate.
	Confirm audiovisual reservations.
	Prepare supplies and training materials for transport to training site.
	Review and rehearse training curriculum.
	Prepare welcome and directional signs for the training site.
	DURING THE WORKSHOP
	Day 1: One hour before start time: Place welcome and directional signs at the facility Set up table to register/sign in participants and distribute training manual, name tags, training handouts and course agenda Check set up of room, check audiovisual equipment, adjust temperature, check supplies (markers, flip charts, etc.)
	Day 1: Conduct training according to agenda Conduct Pre-test.
	Day 2 thru last day, one hour prior to schedule start time: check set up of room check audiovisual equipment adjust temperature Check that all necessary supplies are in place
	Last day: Conduct post-test and evaluation.
	IMMEDIATELY AFTER THE WORKSHOP
	Debrief with co-trainers.
	Send thank you letters to speakers and other key officials.
	Assemble and evaluate pre-and post-test data.
	Analyze workshop evaluation data.
	Prepare report of training.
	1-3 MONTHS AFTER THE WORKSHOP
	Follow up with participants to verify if they were able to apply knowledge and skills obtained during the workshop. Identify additional technical assistance or further training needed.

This page last reviewed: 11/6/2006

Home

Policies and Regulations

Disclaimer

e-Government

FOIA

Centers for Disease Control and Prevention, 1600 Clifton Rd, Atlanta, GA 30333, U.S.A
Tel: (404) 639-3311 / Public Inquiries: (404) 639-3534 / (800) 311-3435

Department of Health
and Human Services