research histology core laboratory...frozen sectioning 1. when the lab receives the sample it will...

Research Histology Core Laboratory

Specialized Services List

1. Immunohistochemistry

Procedures:

1. The investigator must submit a request via the ilabs System. A log sheet (contained within

the request) must be completed. The log sheet must identify the sample name (i.e. name of

slide, block or cassette), description (i.e. murine, porcine, etc.) and sample quantity (number

of blocks, slides, and/or cassettes submitted).

2. The investigator must submit requests in the following manner:

• Antibodies of interest (primary antibody) must be accompanied with the datasheet from

the antibody manufacturer and references for non-established antibodies.

• The minimum amount of antibody to be submitted is 1ml in manufacturer’s vial.

• If available, the investigator must submit positive and negative controls specified by

antibody manufacturer.

• All slides must be positively charged.

3. We recommend developing of all antibodies not included in our panel (see assay

development charges below). All work will be charged according to services rendered.

4. The estimated time of completion for an immunohistochemical request is 14 days. The

investigator will be contacted via email upon completion.

5. The investigator will be responsible for obtaining a pathologist for detailed reading of slides,

or request Pathologist Services from the RHF Core Lab.

6. Immunohistochemical services provided by this lab are intended research purposes only.

The following are charges that will apply to all immunohistochemical staining submitted

to the lab:

Assay development

$468.75/Ab (includes full protocol development and titrations at three dilutions)

Additional titers

$65.00/titration

Manual Immunostain

$35.00/slide

Immunostain only

(Investigator provides antibody)

$25.00/slide (includes specimen, positive and

negative control; sectioning of slide not

included)

Immunostain only

(Core Histology provides antibody)

$31.25/slide (includes specimen, positive and negative control; sectioning of slide not included)

IHC Consultation

$125.00/case(assessment of project requirements)

IHC Preparation Charge $75.00/case (includes the preparation of

user’s accessory reagents)

IHC Research Development

$175.00/case (includes literature or background searches needed to setup assay)

Pathologist’s Services

$120.00/case

If there are any questions regarding the services offered, please phone Jacqueline M.

Furr at X3-0009 or Sherita Meyer-Gauen X2-3119 or via email at

[email protected] or [email protected] , respectively.

mailto:[email protected]


2. RNAase/DNAase-Free Sectioning

Prelimary Requirements: Specimens for frozen sectioning must be submerged in

OCT cryoprotectant, frozen and stored in a -80°C freezer.

Procedures:

Frozen Sectioning

1. When the lab receives the sample it will be stored at -80 degrees Celsius until sectioning.

2. Cryostat and sectioning instruments will be cleaned with disinfectant and treated

with DNAase/RNAase decontaminant.

3. Slides will be printed under routine conditions and will only be handled with gloves

throughout preparation and sectioning.

4. Samples will be equilibrated to -20 degrees Celsius before sectioning begins.

5. Sections will be produced with changes in sectioning area of blade between samples to

prevent cross-contamination.

6. Samples will be stored in lab or investigator provided slide boxes at -80 C until checkout.

7. At checkout, samples should be transported under frozen temperatures after checkout.

Paraffin Sectioning

1. When the lab receives the sample it will be store at room temperature until sectioning.

2. Microtome, sectioning instruments and water bath will be cleaned with disinfectants and

treated with DNAase/RNAase decontaminant.

3. Slides will be printed under routine conditions and will only be handled with gloves

throughout preparation and sectioning.

4. Sections will be produced with changes in sectioning area of blade between samples to


5. Samples will be stored in lab or investigator provided slide boxes at room temperature

until checkout.

Note: This specialized service is also used for LCM sectioning.

3. Mapping

Preliminary Requirements: Samples must be stored at room temperature before

submitting to the lab for sectioning. The

requestor must submit written details for the

embedding arrangement of tissue in paraffin

blocks and the sections on slides.

Procedures:

Embedding

1. Tissue will be placed on the slide according the details submitted. The requestor

can be present for map embedding of tissue.

2. Blocks will be place in queue for sectioning according to the date submitted.

Frozen Sectioning

1. When the lab receives the sample it will be store at room temperature until

sectioning.

2. Cryostat and sectioning instruments will be cleaned with disinfectants and


3. Slides will be printed under routine conditions and will only be handled with

gloves throughout preparation and sectioning.

4. Samples will be stored in lab or investigator provided slide boxes at -80 C until

checkout.

5. At checkout, samples should be transported under frozen temperatures after

checkout.

Paraffin Sectioning

1. When the lab receives the sample it will be store at room temperature until

sectioning.

2. Microtome and sectioning instruments will be washed with disinfectants and


3. Slides will be printed under routine conditions and will only be handled with

gloves throughout preparation and sectioning.

4. Sections will be produced with changes in sectioning area between samples to


5. Samples will be stored in lab or investigator provided slide boxes at room

temperature until checkout.

4. Shavings Preliminary Requirements: Samples must be stored at room temperature if paraffin

blocks or -20 Celsius if frozens, before submitting to the

lab for sectioning.

Procedures: The investigator must submit a request via the ilabs

system for shavings. The requestor must specify if

sections require DNAase/RNAase preparation.

Frozen Sectioning

1. The cutting area will be prepared according to requestor’s protocol requirements

regarding DNAase/RNAase-Free sectioning.

2. Sections will be produced with changes in sectioning area between samples to prevent

cross-contamination of protein extraction.

3. Sections will be produced at great enough thickness to generated curled sections that will

be placed in sterile labeled Eppendorf tubes.

4. Samples will be stored in lab or investigator provided containers at -20 Celsius until

checkout.

5. At checkout, samples should be transported under frozen temperature conditions after

checkout.

Paraffin Sectioning

7. The cutting area will be prepared according to requestor’s protocol requirements

regarding DNAase/RNAase-Free sectioning.

8. Sections will be produced with changes in sectioning area between samples to prevent

cross-contamination of protein extraction.

9. Sections will be produced at great enough thickness to generated curled sections that will

be place in sterile labeled eppendorf tubes.

10. Samples will be stored in lab or investigator provided containers at room temperature

until checkout.

5. Thick Sectioning Preliminary Requirements: Samples must be stored at room temperature if paraffin

blocks or -20 Celsius for frozens before submitting to

the lab for sectioning. The requestor must submit specify

if sections require DNAase/RNAase preparation.

Procedures:

Paraffin Sectioning

1. Cut sections at 8 microns or more according to requester’s protocol requirements.

(Example: 10 microns, 20 microns, 40 microns,etc.)

2. Thick sections will be placed on glass slides.

3. Samples will be stored in slide boxes until checkout.

DNA/ RNA Sectioning

1. Cutting area will be prepared according to requestor’s protocol requirements regarding

DNAase/RNAase-Free sectioning with changes in sectioning area of blade between

samples to prevent cross-contamination of protein extraction.

2. Thick DNA/RNA free sections at 8 microns or more will be placed on glass slides.


LCM Sectioning

1. Cut sections at 8 microns or more according to the requester’s protocol requirements.

2. Thick LCM sections will be placed on regular slides for extraction or slides supplied by

requester.


Frozen Sectioning

1. Cut sections at 8 microns or more in cryostat according to the requester’s protocol

requirements.

2. Thick Frozen sections will be placed on glass slides.

3. Samples will be stored at -20 Celsius in slide boxes until checkout.

4. Samples and slides should be transported in container with dry ice after checkout.

6. Special Stains

Procedures:

1. The specified slides will then be stained using the appropriate requested special stain

procedure.

2. Upon completion of the special stain procedure the slides will be dehydrated and coverslipped

using a mounting medium.

3. Special Stains services provided by this lab are intended for research purposes only.

The following tier system represents special stains offered by the RHCL based on

complexity.

•PAS

•AL. BLUE

•IRON

•CRESYL VIOLET

•WRIGHT STAIN

•TOLUIDINE BLUE

TIER 1

•AFB (ZIEHL NEELSEN)

•GRAM

•VERHOEFFS

•OIL RED O

•MASSON'S

•MUCICARMINE

•CONGO RED

TIER 2

•FITE

•RETICULIN

•MOVAT'S GMS

•FONTANA

•LUXOL FAST BLUE

•ALL OTHER SILVER STAINS

•ALL OTHER COMPLEX STAINS

TIER 3

4. TMA Construction

Procedures:

1. The investigator must submit a request via the ilabs system.

2. The investigator will provide: (a) matching H&E- stained slides and corresponding paraffin

blocks with area(s) to be sampled circled with a marker on the H&E slides, and (b) only a

completed electronic template (via ilabs System) and hard copy of the specimen Array Map

to be used for array construction will be accepted. (See example template below)

• Screening of H&E slides will NOT be performed by Research Histology Core

Laboratory personnel.

• The laboratory will NOT be responsible for requesting slides or paraffin blocks

for microarray tissue projects;

• Blocks and slides will be returned only to the MDACC Investigator.

3. Charges for Tissue MicroArray Block preparation, unstained microarray tissue sections, and

immunohistochemical studies performed on microarray sections are listed below.

4. The estimated time of completion for a Tissue MicroArray request depends on the size of

the project. The investigator will be contacted by email upon completion.

5. Tissue MicroArray services provided by this lab are intended strictly for research


7. The investigator will provide: (a) matching H&E- stained slides and corresponding paraffin

blocks with area(s) to be sampled circled with a marker on the H&E slides, and (b) only a

completed electronic template (via ilabs System) and hard copy of the specimen Array Map to

be used for array construction will be accepted. (See example template below)

• Screening of H&E slides will NOT be performed by Research Histology Core

Laboratory personnel.

• The laboratory will NOT be responsible for requesting slides or paraffin blocks

for microarray tissue projects;

• Blocks and slides will be returned only to the MDACC Investigator.

8. Charges for Tissue MicroArray Block preparation, unstained microarray tissue sections, and

immunohistochemical studies performed on microarray sections are listed below.

9. The estimated time of completion for a Tissue MicroArray request depends on the size of

the project. The investigator will be contacted by email upon completion.

10. Tissue MicroArray services provided by this lab are intended strictly for research.

The following are charges that will apply to all Tissue MicroArray request submitted to the lab:

Service Charge

Annealing (TMA) $15 each

Consultation Fee $125 per session

Manual Construction $943.75per block

Auto Construction $818.75 per block

TMA Cores (1 – 20) $5 per core

TMA Cores (21 – 59) $7 per core

Arranging block and slides $87.50 per block

Microscopic ID $70 slide

Serial Sections $12.50 per slide

Unstained Section $6.25 per slide

TMA H&E Only $3.50 per slide

H&E stained array slides $7.00 per slide

Special stains on TMA slides Tier I - $7.50 per slide / Tier II - $13.50 per slide

IHC performed on tissue array slides $28.00 per slide

EXAMPLE

RHF Histology Core Lab TMA Template

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15

1

2

3

4

5

6

7

8

9

10

If there are any questions regarding the services offered, please phone Jacqueline M. Furr at X3-0009 or

Sharon LeBlanc X2- 3119 or via email at [email protected] or [email protected] respectively.



5. RHCL Pathologist’s Review Procedures:

A. Slide Scanning

1. The investigator must submit a request via the iLabs Chargeback System. A log sheet

(contained within the request) must be completed. The log sheet must identify the

sample name (i.e. name of slide, block or cassette), description (i.e. tumor, A1, etc.) and

sample quantity (number of blocks, slides, and/or cassettes submitted).

2. The slides must be submitted in the following manner:

• All slides must be dry and free of excess mounting media.

• Slides that have air bubbles or the mounting media has dried out must be re-coverslipped

prior to bringing slides for scanning.

• Broken, chipped, or repaired slides cannot be scanned.

• All slides will be routinely scanned at 20X magnification unless otherwise requested.

• Necrotic tissue, poorly stained slides, and slides without counterstain take longer to scan,

and will not be of standard quality.

• Scanned slides will be stored for 30 days. Scanned slides can be downloaded onto a hard

drive provided by the requester.

• The estimated time of completion for slide scanning requests is 5-8 days. The

investigator will be contacted via iLabs notification upon completion.

Slide Scanning Fee:

B. Pathologist’s Review


2. Stained H&E slides must be submitted with request for Pathologist Review when the

detection of proper staining or the presence of tumor are needed.

3. IHC consultations with the Pathologist are scheduled by appointment. The consultations

are for discussing antibody selection, optimization of antibodies and IHC protocols with

the investigators.

Services provided:

Prepared Slides (H&E) 2 or more days $120.00 per case Review of stained slides to acknowledge effective staining or the presence of tumor.

IHC Consultation By appointmen t $120.00 Consultations are held for 1 hour. Every 30 minutes after initial hour is $10.00.This service includes antibody

selection, optimization of antibody and IHC protocols.

The RHCL Pathology review is not provided for diagnosis or a case analysis

Quantity Cost

Slide Scan (1-50 slides) $8.00 per slide



Slide Scan (501 slide and up) $5.00 per slide

research histology core laboratory...frozen sectioning 1. when the lab receives the sample it will...

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