The Georgia Society for Histotechnology · Page 10 Spring 2009 The Georgia Society for...

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Spring 2009

The Geor g ia Soc ie ty fo r H i s t o te chno logy

MICROTIME MICROTIME MICROTIME

Presidents Message Page 2

Board of Directors Page 3

Characteristics and Types of Fixatives Page 5-7

Schedule at Glance—NSH Page 9-11

Letter from GSH Secretary Page 12

GSH Membership Form Page 15

NSH Membership Form Page 17

Inside…..

Issue 6 FALL 2011

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President’s Message Fall 2011

It seems like the summer just melted away with all the heat. I hope each of you had a wonderful and

restful summer with your families. NSH is just around the corner and for those who will be going I hope you

have a good trip and learn about all the new technology. In the journals I still receive, and yes I read them too, it

seems as if new test and procedures are being introduced daily. It is hard to keep up and ahead of the game.

I hope you have made your reservations for Region III at Callaway Gardens and if you have not please

make your reservations soon. The phone number is 1-800-225-5292 and the group code is 78K711 with a room

rate is $109. They will not bill your credit card, per our contract, until the day of check in. It is very impor-

tant for us to have these rooms booked soon to ensure that no one gets left out. We only blocked a certain num-

ber, but they will honor the rates until Callaway runs out of rooms.

This is a big meeting, with a great lineup of workshops and speakers. I want to compliment Carl and

the Education committee for some awesome work in obtaining a virtual Who’s Who in speakers. We are ex-

pecting a large number of Vendor’s as well. Wanda Simons is working with them and they are already excited

about coming to the meeting.

We will be releasing the program very shortly and then registration can begin. There will be some ex-

citing things occuring during the meeting and of course if you bring your family, room reservations will get you

into the gardens for free. The flowers should be bursting forth by this time of the year.

I hope each of you have a great fall and holiday season. If any of the Board or Officers can be of assis-

tance don’t ever hesitate to call or email us. We will respond to your needs.

Mike Ayers

President GSH

[email protected]

678-378-4663

President Letter….

mailto:[email protected]

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BOARD OF DIRECTORS—2011 GSH PRESIDENT

Mike Ayers

Newnan, GA

PH: 770- 253-7462 - Cell: 678-378-4663

FX: 678-378-4663

Email: [email protected]

GSH VICE PRESIDENT & GSH EXHIBIT LIAISON:

Wanda Simons HT, (ASCP)

Snellville, GA 30078

PH: (770)-736-2565 FX: (770)-736-2368


GSH TREASURER & BUDGET AND FINANCE CHAIR

Mary Anne Taylor

Macon, GA 31216

PH: 478-788-1286

FAX: 478-788-1286


GSH SECRETARY & WEB MANAGER

Shirley A. Powell HT(ASCP)HTL, QIHC

Macon, GA 31207

PH: 478-301-2374

FX: 478-301-5489


GSH MEMBERSHIP CHAIR

Nancy Crane

Lawrenceville, GA 30046

PH: 678-312-4506

PH: 770-963-8958


GSH AWARDS CHAIR

Terri Brown

Lilburn, GA 30047

PH: 404-845-5423

FX: 404-303-3706


Connie Micko

Stone Mountain, GA 30088

PH: 770-413-2623


GSH BYLAWS & LEGISLATIVE CHAIR

Larry Fields

PH: (803) 917-4536

Email:[email protected]

GSH Ed Committee Speaker Liaison

Donna Smith

Lawrenceville, GA 30046

PH: 678-312-4206

FX: 770-682-2219

PH: 404-717-6535 (cell)


MICROTIME EDITOR

Carole Fields

Marietta, GA 30067

PH: 404-895-9604

Email: [email protected] Email: [email protected]

GSH EDUCATION CHAIR – PROGRAM COORDINATOR

Carl Sagasser

Albany, GA 31707

PH: 229-317-6974

FX: 229-317-6682


GSH SYMPOSIUM REGISTRAR

Harriet Baker

Griffin, GA 30224

PH: 770-228-2721

PH: 770-229-1272


GSH HISTORIAN

Janet Hobbs

Augusta, GA 30912

PH: 706-721-8515



GSH PUBLIC RELATIONS

Fran Lee Adams Davis

PH: 770-991-8000 5408

FX: 770-991-8689

McDonough, GA 30252

PH : 770-689-6484


GSH Ed Committee Program Designer

Michael Bourgeois

Smyrna, GA 30082

PH: 404-213-8887


GSH CAREER COMMITTEE

Angie Rawl

VA Medical Center

PH: 706-733-0188 EXT 2957

FX: 706-823-3965


PH: 803-564-6128

FX: None




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Fixation Part 1: Characteristics and Types of Fixatives

by René J. Buesa B.Sc. HTL (ASCP) (Ret.) [ [email protected] ]

Fixation is by far the most important step in tissue processing because it will not only pre-vent the decay of cell and tissue structures, but will also preserve the structure of tissues permitting a proper infiltration with the melted paraffin. Good infiltration and the subsequent ability to prepare sections of high quality of the processed tissues depend fundamentally on good fixation. A poorly infiltrated tissue consequence of poor fixation is an irreversible problem, no matter what you try to do after the fact. All those “post-fixation” remedies that many of us have used trying to save a poorly fixed specimen are essentially worthless no matter what anecdotal experiences one may have.

Besides the fixative itself, which should be adequate to the specimen to be fixed (Table 1), there are other factors affecting the process amongst of which are volume of fixative to specimen volume ratio, temperature and pH, agitation in the fixative and, most importantly, time in the fixative. It is also necessary to point out that fixatives can be simple or complex, made up of just one or sev-eral chemicals.

One can preserve tissues by deep freezing them although this is not a fixation method. On the other hand, heat fixing is a standard procedure in microbiology where smears are heat fixed before being stained. With regards to conventional liquid fixatives for tissues, temperatures above room temperature (RT) determine a faster fixation of tissues. In the case of formalin the tissues will half-bind in 11 hours at 25ºC RT, but will reach the same binding level in only 5.5 hours at 37ºC RT (1).

Recently the use of microwave ovens (MW) for tissue fixation is becoming more frequent but it is important to stress that the rapid fixation is not caused by the microwaves, but by the tem-perature increase they cause (2).

There are literally hundreds of fixatives most developed between 1850 and 1950 (3) and each have its own peculiarities and belong to groups with overall similar characteristics in a way that there are oxidizing, protein denaturing and cross linking agents, coagulating fixatives and even some used since time immemorial, like mercury chloride and picric acid, whose fixation mecha-nisms are still not well understood. It is also worth noting that many of the chemicals used in many fixing solutions are highly toxic and now forbidden by OSHA.

Of all the fixatives used, the “gold standard” against which all new fixatives and substitutes are compared, is the neutral buffered formalin (NBF) that is still the fixative of choice of the majority of US histolabs or in the totality in other countries, like Mexico and Russia. Before starting to de-scribe the peculiarities of tissue fixing with NBF, it is important to understand other issues.


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When reading any histology material the first thing that is evident is the absolute inconsistency regarding the ratio fixative volume to tissue volume. The “rule of thumb” for many is 20:1 (20 times the fixative vol-ume to the tissue volume) but this is not a standard. Out of 100 references dealing with this issue; 4 rec-ommend a ratio of less than 10:1; 50 recommend a ratio from 10 to less than 20:1; 39 recommend ratios of 15 to 20:1 and 7 recommend ratios of more than 20:1. Of course that all these references report good fixations results meaning that the volume of fixative to tissue is a less than settled issue.

Another very important issue in fixation, especially important for complex fixatives, is that each chemical component will penetrate the tissue at a specific rate, regardless of the mixture they are in. This penetration rate is called diffusion coefficient, expressed as “k” in the equation [d = k √ t ], where “d” is the penetration of the fixative into the tissue in millimeters, and “t” is the time in hours the tissue is exposed to the fixative.

When dealing with complex fixatives the tissue will be fixed at different rates by each component. For example: the fixative Formalin-Alcohol-Acetic (FAA), widely used for plant materials and sometimes for human materials, is composed of 10 parts of pure (37-40%) formalin, 50 parts of pure (100%) ethanol, 5 parts of pure (“glacial”) acetic acid, and 35 parts of distilled water.

Each of these components have a different diffusion coefficient with values of k = 0.78 for formalin, k = 1.0 for ethanol, and k = 1.2 for acetic acid. This means that the acetic acid will reach the cells before the other two components and formalin will be the last. This staged fixation process will produce a final tissue image different to that obtained with formalin alone, and why it is that different fixatives and their mixtures produce different final microscopic images. Consequently, when developing a formula for a new fixative it is important to take into account their diffusion coefficients.

In Table 1 there are some examples of different subjects and the fixatives used for each and it is worth noting that NBF is considered amongst the fixatives for eight of the twenty-two subjects included, and the general fixative of choice.

Fixation with formalin is a very peculiar process with three different stages: penetration, binding, and cross-linking all happening simultaneously but a very different rate, cross-linking being the final objec-tive.

Essentially fixation with formalin is a very slow process and will be the subject of the next Note.

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Table 1 - Fixatives of choice for different subjects (3)

Subject Fixative

Amoebas Schaudin’s (contain mercury chloride)

Carbohydrates Alcohol-acetone (3 pure ethanol + 1 acetone); FAA

DNA and RNA Carnoy’s; 70% ethanol

Enzymes Zenker’s (contains mercury chloride)

Fribrin Mercuric chloride; Bouin’s; NBF

Glycogen Pure ethanol; Gendre’s

Iron pigments Alcoholic formalin

Lipases Cold acetone

Lipids and lipoids Osmium tetroxide; Aoyama’s; NBF

Lymph nodes B5 (contains mercury chloride); NBF

Muco substances Formalin saline (with sodium chloride)

Nuclear detail Carnoy’s (6 pure ethanol + 3 chloroform +1 acetic acid)

Nucleic acids Methacarnoy (methanol instead of ethanol)

Overall specimens NBF

Phosphatase Cold acetone; NBF

Phospholipids Helly’s; Zenker’s (both contain mercury chloride)

Polysaccharides Cold alcohol-formalin; Gendre’s; Helly’s; Rossman’s

Red blood cells NBF; Carnoy’s

Silver stains NBF; Helly’s; Zenker’s

Smears (blood) Methanol (after smear is air dried)

Smears (body fluids) Clarke’s (3 pure ethanol + 1 acetic acid)

Weigert’s elastic stain NBF

Fixatives containing mercury chloride have been almost totally eliminated because of safety concerns].

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[References:

1 - Buesa, RJ: Histology without formalin? Ann.Diag.Pathol., 2008; 12(6):387-3962 - Leonard JB, Shepardson SP: A comparison of heating modes in rapid fixation techniques for electron microscopy. J.Histochem.Cytochem., 1994; 42(3):383-391

3 - Buesa RJ: Histology Review. Science & Business Institute, North Miami Beach, 1996; pp. 141

2012 is Election Year for GSH!

Think about who you might want to nominate for office in your

society. We have 155 members and we will be giving an award to

the person who recruits the most members. This is your society…

It takes all of us to make it GREAT! It is your profession and you

are the ones to support GSH and to make it grow! Membership is

free. Send in your membership NOW! You will be glad you did.

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When Symposium Dates: September 16-21, 2011

Scientific Exhibits: September 18-20, 2011

Where

Sessions & Scientific Exhibits will take place in the Duke Energy Convention Center,

525 Elm Street, Cincinnati, OH, 45202.

NSH has reserved rooms at the Hyatt Regency, Millennium Hotel and the Hilton Cincinnati

See you there!

Scott Bryant

www.sbryantinc.com

Mobile: 404-697-9590

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Schedule at a GlanceSchedule at a GlanceSchedule at a Glance updated May 4, 2011

Friday, September 16, 2011 11:00am - 6:00pm Registration Opens

4:00pm – 5:00pm Committee Chairs Meeting

5:00pm – 6:30pm Region Director/President’s Council

6:30pm – 7:00pm Mentors Meet & Greet

7:00pm - 8:00pm First Time Attendee Welcome Reception

Saturday, September 17, 2011 7:00am - 5:00pm Registration

7:00am - 8:00am Continental Breakfast Available

7:00am – 7:45am NSH Committee Meetings

• Budget & Finance

• Education/CEU

• Membership

• Quality Control

• VIR

8:00am - 11:30am Workshops 1-13

11:30am - 1:00pm Lunch on Your Own

12:00pm - 12:45pm Resource Group Meetings:

• Instructors of Histotechnology

• Journal Editorial Board

• State Newsletter Editors

• State Meeting Planners

1:00pm - 2:30pm Workshops 14-24

3:00pm – 4:30pm Workshops 25-36

4:35pm - 5:30pm NSH Region Meetings (1-9)

4:35pm - 5:30pm International Attendee Meeting

Sunday, September 18, 2011

7:00am - 5:00pm Registration


7:00am – 7:45am NSH Committee Meetings

• Hard Tissue

• IHC

• Health & Safety

• Legislative

• PR


9:45am – 10:45am Keynote Lecture

10:45am - 4:30pm Exhibits Open for Attendees

10:45am - 12:15pm Poster Sessions


7:00pm – 11:30pm NSH Awards Banquet

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Monday, September 19, 2011



8:00am - 4:00pm Board of Directors Meeting


9:45am – 10:45am Keynote Lecture

9:00am - 3:30pm Exhibits Open for Attendees

11:30am - 1:30pm Lunch On Your Own


6:00pm Charter Member Reception

Tuesday, September 20, 2011




9:00am - 1:00pm Last Day of Exhibits

11:30am - 1:00pm Lunch on Your Own

12:30pm - 4:00pm Workshops 96-105

4:45pm - 6:00pm NSH Membership Meeting

6:00pm - 7:00pm Committee Meetings:

• Bylaws

• Nominations/Election

7:00pm NSH Night at the Ballpark – Cincinnati Reds vs Houston Astros

Wednesday, September 21, 2011



8:00am - 11:30am Workshops 106-113

11:30am - 1:00pm Lunch On Your Own

1:00pm - 4:30pm Workshops 114-121

6:00pm Sign In for House of Delegates

7:00pm House of Delegates Convenes

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Hi GSH members,

Just wanted to let you know we have a great lineup of speakers for the Georgia Society for Histotechnology

meeting in 2012 when we are hosting the Region III. It will be at Callaway Gardens, the Mountain Creek

Inn in Pine Mountain GA April 13-15th. The program will be finalized soon and up on our website at

www.histosearch.com/gsh<http://www.histosearch.com/gsh> very soon. The flyer with the hotel information

is already there except for the GSH group #. I urge you to make your reservations now, the sooner the

better, at 1-800-225-5292. Spring is a popular time at Callaway and the rooms fill up fast. Be sure to use the

GSH group # 78K711 to get the discounted room rate of $109. The price of your room will get you access to

the gardens. Your credit card will NOT be billed until you register at the hotel in April 2012. There are only

a certain number of rooms blocked so don't delay.

Our speaker lineup to date is below. This is a great way to get your CEUs and also to visit a wonderful south-

ern jewel. There is something for everyone and April will be a gorgeous month for our meeting. The gardens

are in bloom, the golf courses are outstanding and there is a beach for the kids. The butterfly pavilion is al-

ways a big hit with everyone.

To date our speakers and topics are:

Taiquanda Winbush- HT/HTL Review

Adrianna Eaton - Antibody Challenge

Claudia Lawson - Advances in Molecular Testing

Joyce Weems - CPT Coding - Sweet Dream or Nightmare?

Marvin Hanna - The Molecular Pathology Lab of the Future

Ada Feldman - Troubleshooting Hematoxylin and Eosin Staining

Wanda Jones - IHC Controls---The Good, The Bad and The Ugly!

Ada Feldman - The Joy of Histology or What We Can Learn from the Kitchen

Steven Westra - Get the most out of your Immunohistochemistry: A Balance between Convenience, Cost, Education

and Flexibility

Lamar Jones - Real Time Rapid Tissue Processing

Ely Klar - Human Tissues: Histological Identification of the Different Types

So come to Georgia in April 2012, it is going to be a Histo-terrific Meeting.

Shirley A. Powell, HT(ASCP)HTL, QIHC

GSH Secretary

Please share with others that may not be members!

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Georgia Rep...

Sharon Wehman [email protected]

678-462-6349

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GEORGIA SOCIETY FOR HISTOTECHNOLOGY

MEMBERSHIP OR RENEWAL FORM

Please fill out all blanks COMPLETELY - PLEASE PRINT and Mail to:

NANCY CRANE

2366 CELBA COURT

LAWRENCEVILLE, GA 30043

Name _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _

Home City _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ ST _ _ _ _ _ _ _ _ _ _ _ Zip _ _ _ _ _ _ _ _ _ _ _ _ _ _ _

Home Phone _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Fax _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _

Home Email _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _

Employer _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _

Work Address _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _

Work City _ _ _ _ _ _ _ _ _ _ _ _ _ State _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Zip _ _ _ _ _ _ _ _ _ _ _ _ _ _ _

Work Phone _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ Fax _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _

Work Email _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _

NSH MEMBER _ _ _ _ _ _ Y ES _ _ _ _ _ _ _ NO

GSH recommends that you also consider joining NSH in order to be eligible to serve as a representative of GSH in the House of Dele-

gates at the annual Symposium and actively participate in your society. Only NSH members who are GSH members can serve in the

HOD.

*******Date: _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ for year 2011 ****** REQUIRED

Membership is free to anyone working in histology in Georgia but you must fill out a form each year to be a member. If you do not live

in but work in Georgia, you qualify. If you do not work in Georgia you may still be a non-voting member with other member privileges

such as discount rates at meetings.

MEMBERSHIP YEAR IS FROM JANUARY TO DECEMBER

Are you interested in serving on a committee? _ _ _ _ _ _ _ Do you prefer mail sent to: Home? _ _ _ Work? _ _ _ _ _ _ _

It is very important to notify us of changes in your information as soon as possible so we can keep you informed.

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Georgia Rep…..

Sandy Schmitz

404-697-5262

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TO:

Carole Fields, HT (ASCP)

962 Hickory Leaf Ct.

Marietta, GA 30065

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