Reticulum

Controls Slides Histopathology Reticulum

Validation Stain: Gordon-Sweets
Other Applicable Stains: Snook Reticulun and Gomori Stain for Reticular Fibers

 

Tissue: 
(+) Liver
Adhesive or Charged Slides: 
Superfrost +

Product Options:

Part # 4620A10/Set $46.00
Part # 4620B98/Set $330.00

Reticulum Control Slides contain a section of positive staining liver.

                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                               

PRODUCT DESCRIPTION:

The enclosed positive control slides are intended to be used to verify histological techniques and reagent reactivity.  These slides are to be used for the qualitative purpose of determining positive or negative results, and are not intended to be used for any quantitative purpose.  The first serial section within the control box is stained and provided for your reference.  Before using the unstained slides, review the enclosed stained slide with your pathologist to ensure that this tissue source is acceptable.  Newcomer Supply will not accept a return with missing slides in the series. Newcomer Supply guarantees reactivity of these control slides for one year from the date of receipt.  Revalidate after one year to verify continued reactivity.  Store at 15-30°C in a light deprived and humidity controlled environment.

These positive control slides were produced from human surgical or autopsy tissues under carefully controlled conditions.  Reasonable measures are used to deliver quality control slides that are as consistent as possible.  However, characteristics of quality control slides may be dissimilar due to variations in the reagents, stains, techniques, laboratory conditions, and tissue sources used. Newcomer Supply Laboratory uses a manual method of performing quality control procedures, specifically avoiding automation, in order to provide reactive control slides for even less aggressive methods of staining that our customers may be using.

 

CONTROL SLIDE VALIDATION:

With Reticulum, Gordon and SweetsStain Kit: Part 9168A/B Individual Stain Solution
Solution A: Potassium Permanganate 1%, Aqueous 250/500 ml Part 13393
Solution B: Oxalic Acid 1%, Aqueous 250/500 ml  
Solution C: Ferric Ammonium Sulfate 2.5%, Aqueous 250/500 ml  
Solution D: Silver Nitrate 10%, Aqueous 50/100 ml Part 13806
Solution E: Ammonium Hydroxide 28-30%, ACS  50/100 ml Part 1006
Solution F: Sodium Hydroxide 3%, Aqueous 50/100 ml  
Solution G: Formalin 10%, Aqueous 250/500 ml  
Solution H: Gold Chloride 0.2%, Aqueous 250/500 ml Part 11286
Solution I: Sodium Thiosulfate 5%, Aqueous 250/500 ml Part 1389
Solution J: Nuclear Fast Red Stain, Kernechtrot 250/500 ml Part 1255

For storage requirements and expiration date refer to individual product labels.    

 

APPLICATION:

Newcomer Supply Reticulum Control Slides are for the positive histochemical staining of reticulum fibers; regarded as specialized connective tissue fibers. 

 

METHOD:

Fixation: Formalin 10%, Phosphate Buffered (Part 1090)

Technique:  Paraffin sections cut at 5 microns on Superfrost® Plus

Solutions:  All solutions manufactured by Newcomer Supply, Inc.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. All glassware/plasticware must be acid cleaned prior to use.
    1.  See Procedure Notes #1 and #2.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1.  See Procedure Notes #3 and #4.
  3. Oxidize sections in Solution A: Potassium Permanganate 1%, Aqueous for 5 minutes.
  4. Wash in running tap water for 2 minutes; rinse in distilled water.
  5. Bleach in Solution B: Oxalic Acid 1%, Aqueous for 2 minutes or until sections are colorless.
  6. Wash in running tap water for 2 minutes; rinse in distilled water.
  7. Sensitize sections in Solution C: Ferric Ammonium Sulfate 2.5%, Aqueous between 15 to 20 minutes.
  8. Rinse slides in several changes of distilled water.
  1. Prepare Ammoniacal Silver Working Solution:
    1.   Place 5 ml of Solution D: Silver Nitrate 10%, Aqueous in a flask; add Solution E: Ammonium Hydroxide 28-30%, ACS drop by drop, swirling continuously until formed precipitate completely dissolves.  Do not add any excess Ammonium Hydroxide. 
    2.   Add 5 ml of Solution F: Sodium Hydroxide 3%. Aqueous.
    3.   Re-dissolve formed precipitate with Solution E: Ammonium Hydroxide 28-30%, ACS until a faint cloudiness remains.
    4. If proceeded too far and no cloudiness remains, add Solution D: Silver Nitrate 10%, Aqueous, drop by drop, until one drop causes the solution to become permanently cloudy.  A faint cloudiness is the optimum.
    5.   Bring solution total volume to 50 ml with distilled water; filter.
  2. Impregnate sections in filtered Ammoniacal Silver Working Solution for 2 minutes.           
  3. Rinse slides well in running distilled water for 1 minute.
    1.  See Procedure Note #5.
  4. Reduce sections in Solution G: Formalin 10%, Aqueous for 2 minutes.
  5. Rinse in running tap water for 3 minutes.
  6. Tone in Solution H: Gold Chloride 0.2%, Aqueous for 10 minutes.
  7. Rinse well in distilled water.
  8. Place slides in Solution I: Sodium Thiosulfate 5%, Aqueous for 1 minute.
  9. Wash well in tap water for 2 minutes; rinse in distilled water.
  10. Counterstain with Solution J: Nuclear Fast Red Stain, Kernechtrot for 5 minutes.
    1.  Shake solution well before use; do not filter.
  11. Rinse well in distilled water.
    1. See Procedure Note # 6.
  12. Quickly dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Reticulum fibers Black
Background Red

 

PROCEDURE NOTES:

  1. Acid clean all glassware/plasticware (Part 12086) and rinse thoroughly in several changes of distilled water.  Cleaning glassware with bleach is not equivalent to acid washing.
  2. Plastic (Part 5500, 5501), plastic-tipped or paraffin coated metal forceps must be used with silver solutions to prevent precipitation of silver salts.  No metals of any kind should come in contact with silver solutions.
  3. Drain staining rack/slides after each step to prevent solution carry over.
  4. Do not allow sections to dry out at any point during staining procedure.            
  5. This rinse step is critical for good reticulum demonstration.  If rinsing is insufficient, excessive background staining may occur.
  6. Wash well after Nuclear Fast Red Stain, Kernechtrot to avoid cloudiness in dehydration steps. 
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Carson, Freida L., and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 177-179.
  2. Bancroft, John D., and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 155-156.
  3. Gordon, Harold, and Henry Sweets. "A Simple Method for the Silver Impregnation of Reticulum." American Journal of Pathology 12.4 (1936): 545-552.
  4. Modifications developed by Newcomer Supply Laboratory.