STEINER STAIN KIT PROCEDURE [PDF]

PRINCIPLE: This kit demonstrates Fungi, Helicobacter pylori, Legionella ... Place slide in 0.1% SILVER NITRATE for 5 min

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Idea Transcript


STEINER STAIN KIT PROCEDURE Item# KT035 KIT KIT INCLUDED:

100ml. 100ml. 1.5gm.

OXIDIZER SOLUTION OXIDIZER SOLUTION HYDROQUINONE

100ml. 100ml. 5ml.

10% ZINC FORMALIN 10% ZINC FORMALIN 1% SILVER NITRATE

PRINCIPLE:

This kit demonstrates Fungi, Helicobacter pylori, Legionella pneumophila, and Spirochetes.

SPECIMEN:

Any well fixed paraffin embedded tissue cut at 3 to 4 microns.

QUALITY CONTROL:

Recommended Control Slide: Fungus, Helicobacter pylori, Legionnaires, Spirochete.

SOLUTION: Reducing Solution: Prepare at Step #10 1% Hydroquinone (Make Fresh) ................ 25ml 2.5% Gum Mastic...................................... 15ml 1% Silver Nitrate ....................................... 6 drops Prepare by adding 0.25 gm Hydroquinone to 25 ml Distilled water and mix until completely dissolved. Add 15ml of 2.5% Gum Mastic, mix thoroughly, and filter solution through medium filter paper. Add 6 drops of 1% Silver Nitrate and mix thoroughly. PROCEDURE: i. Preheat water bath to 70°C. ii. Deparaffinize slide using Xylene or a Xylene Substitute and hydrate through alcohols to Tap water. iii. Place slide in OXIDIZER SOLUTION for 20 minutes. iv. Rinse slide through 2 changes of Distilled water. v. Place slide in 10% ZINC FORMALIN for 5 minutes. (At this time place 20 ml of 0.1% SILVER NITRATE in the water bath.) vi. Rinse slide through 2 changes of Distilled water. vii. Place slide in 0.1% SILVER NITRATE for 5 minutes at 70°C; discard solution after this step. viii. Rinse slide through 2 changes of Distilled water. ix. Dehydrate slide through 1 change of fresh Absolute Alcohol. x. Place slide in 2.5% GUM MASTIC for 3 minutes. (At this time, prepare Reducing Solution and place in the water bath.) xi. Air dry slide for 1 minute or until the gum mastic is completely dry. xii. Place slide in Reducing Solution at 70º C for 10 to 15 minutes or until section is tan to brown; discard Reducing Solution after this step. xiii. Dehydrate slide through 3 changes of fresh Absolute Alcohol. xiv. Clear slide through 3 changes of fresh Xylene or Xylene Substitute. xv. Coverslip slide immediately using a permanent mounting media. RESULTS: Cat Scratch Fever, Legionella pneumophila, DARK BROWN, GRAY, OR Spirochetes, Helicobacter pylori, and Fungi: BLACK Background: YELLOW, TAN, OR BROWN REFERENCE: Elias JM, Green C: Modified Steiner method for the demonstration of Spirochetes in tissue, Am J Clin Pathol; 1979, 71: 109 - 111. For Research Use Only. Not for Diagnostic or Therapeutic Work. DBS will not be held responsible for patent infringement or other violation that may occur with the use of our product.

DBS 1020 Serpentine Lane, # 111, Pleasanton, CA 94566 Tel: 925 484 3350, Fax: 925 484 3390

STEINER STAIN KIT MICROWAVE PROCEDURE KIT COMPONENTS INCLUDED:

100ml. OXIDIZER SOLUTION 100ml. 0.1% SILVER NITRATE 1.5gm. HYDROQUINONE

100ml. 10% ZINC FORMALIN 125ml. 2.5% GUM MASTIC 5ml. 1% SILVER NITRATE

PRINCIPLE:

This kit demonstrates Fungi, Helicobacter pylori, Legionella pneumophila and Spirochetes.

SPECIMEN:

Any well fixed paraffin embedded tissue cut at 3 to 4 microns.

QUALITY CONTROL: SOLUTION:

Recommended Control Slide: Fungus, Helicobacter pylori, Legonnairs, Spirochete.

REDUCING SOLUTION: Prepare at Step #10 1% Hydroquinone (Make Fresh) ................ 25ml 2.5% GUM MASTIC ................................... 15ml 1% SILVER NITRATE ............................... 6 drops Prepare by adding 0.25gm HYDROQUINONE to 25ml Distilled water and mix until completely dissolved. Add 15ml of 2.5% GUM MASTIC, mix thoroughly, and filter solution through medium filter paper. Add 6 drops of 1% SLVER NITRATE and mix thoroughly. PROCEDURE: i. Deparaffinize slide using Xylene or a Xylene Substitute and hydrate through alcohols to Tap water. ii. Place slide in OXIDIZER SOLUTION for 20 minutes. iii. Rinse through two changes of Distilled water. iv. Place slide in 10% ZINC FORMALIN for 5 minutes. v. Rinse slide through two changes of Distilled water and hold in Distilled water until Step #7. vi. In a loosely capped Slide Jar (available from A.M.T.S.) heat 20 ml of 0.1% SILVER NITRATE in a microwave oven for 10 seconds at full power. Repeat as needed until solution is hot, but do not allow solution to boil. Remove Slide Jar from microwave, tighten Slide Jar cap and agitate to equalize solution temperature. vii. Place slide into the hot 0.1% SILVER NITRATE, agitate thoroughly and incubate for 2 minutes. viii. Rinse slide through 2 changes of Distilled water. ix. Dehydrate slide through 2 changes of fresh Absolute Alcohol. x. Place slide in 2.5% GUM MASTIC for 3 minutes. (During this step, prepare Reducing Solution.) xi. Air dry slide for 1 minute or until the gum mastic is completely dry. xii. In a loosely capped Slide Jar heat 20 ml of Reducing Solution in a microwave oven for 25 seconds at full power. Tightly cap Slide Jar, invert several times and place slide into the Reducing Solution. Loosely cap Slide Jar, return Slide Jar to the microwave and heat at full power until the Reducing Solution is hot, but do not allow solution to boil. Incubate slide for 3 minutes, and then heat again at full power until the Reducing Solution is hot. Incubate slide for 2 to 3 minutes; section color should be tan to light brown. xiii. Rinse slide quickly in Distilled water. xiv. Dehydrate slide through 3 changes of fresh Absolute Alcohol. xv. Clear slide through 3 changes of fresh Xylene or Xylene Substitute. xvi. Coverslip slide immediately using a permanent mounting media. RESULTS: Cat Scratch Fever, Legionella pneumophilia, DARK BROWN, GRAY, OR Spirochetes, Helicobacter pylori, and Fungi: BLACK Background: YELLOW, TAN, OR BROWN REFERENCE: Elias JM, Green C: Modified Steiner method for the demonstration of Spirochetes in tissue, Am J Clin Pathol; 1979, 71: 109 - 111. For Research Use Only. Not for Diagnostic or Therapeutic Work. DBS will not be held responsible for patent infringement or other violation that may occur with the use of our product.

DBS 1020 Serpentine Lane, # 111, Pleasanton, CA 94566 Tel: 925 484 3350, Fax: 925 484 3390

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