EP0345295A1 - Reagent and method for detecting cells with phagocytic activity - Google Patents

Reagent and method for detecting cells with phagocytic activity

Info

Publication number
EP0345295A1
EP0345295A1 EP19880902576 EP88902576A EP0345295A1 EP 0345295 A1 EP0345295 A1 EP 0345295A1 EP 19880902576 EP19880902576 EP 19880902576 EP 88902576 A EP88902576 A EP 88902576A EP 0345295 A1 EP0345295 A1 EP 0345295A1
Authority
EP
European Patent Office
Prior art keywords
solution
weight
reagent
neutral red
phagocytic activity
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP19880902576
Other languages
German (de)
French (fr)
Inventor
Katalin Polgar
György ABEL
Ilona Harsanyi
Lászlo NAGY
Sándor SIPKA
Edit Vaszari
Ivan Daroczi
Zoltán PAPP
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Reanal Finomvegyszergyar Rt
Original Assignee
Reanal Finomvegyszergyar Rt
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Reanal Finomvegyszergyar Rt filed Critical Reanal Finomvegyszergyar Rt
Publication of EP0345295A1 publication Critical patent/EP0345295A1/en
Withdrawn legal-status Critical Current

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5094—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for blood cell populations
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing

Definitions

  • the invention relates to a reagent and a method for detecting cells with phagocytic activity.
  • the solution of neutral red in distilled water is of acidic character (the pH of a solution containing 1 mg/ml of neutral red is 3.1), and, owing to its acidity and hypoosmotic effect, this solution strongly damages the cells. None of the aqueous buffer and nutrient solutions utilized in routine clinical practice proved to be appropriate to dissolve neutral red, since after dissolution precipitation occurs. Organic solvents appeared to be less suitable; thus e.g. neutral red in ethanol solution does not give appropriate staining, whereas the dimethyl sulphoxide solution is difficult to sterilize.
  • the invention aims at providing a reagent solution and diagnostic method for detecting cells with phagocytic activity which is easy to apply and gives highly reliable results.
  • a neutral red-containing cytologic reagent solution should be approximately isoosmotic in relation to the potentially pathologic cells. It is also essential that the mixture of the reagent solu tion formed with the sample to be examined (amniotic fluid) should be almost neutral. It has also been observed that lysozyme enzyme significantly increases the stain uptake of the cells, moreover it Increases the stability of the reagent solution, too. Finally it has been found that the reagent solution should contain a saccharide as an energy source for the cells. When a reagent solution storable for a prolonged period of time is to be prepared, preferably a preservative should also be added to the mixture.
  • the invention relates to a neutral red-containing reagent solution for detecting cells with phagocytic activity.
  • the reagent solution according to the invention comprises 0.2-0.8 % by weight of neutral red, 0.1-0.25% by weight of lysozyme, 0.8-2.0 % by weight of sodium chloride, 0.1-0.3 % by weight of a saccharide and optionally 0.02-0.6 % by weight of a preservative in an aqueous solution with a pH of 3.0 to 8.0, preferably 3.5 to 5.0.
  • the invention also relates to a method for detecting cells with phagocytic activity by staining them with neutral red. According to the invention one proceeds as follows: 0.2-0.8 % by weight of neutral red, 0.1-0.25 % by weight of lysozyme, 0.8-2.0 % by weight of sodium chloride, 0.1-0.3 % by weight of a saccharide (preferably glucose) and optionally 0.02-0.6 % by weight of a preservative are dissolved in water, the pH of the solution is adjusted to 3.0-8.0, preferably 3.5-5.0, the solution is sterilized by filtration, thereafter one part by volume of the resulting reagent solution is admixed with 3-8 parts by volume of a sample to be examined, preferably amniotic fluid, and the resulting cell suspension is examined in a haemocytometer in a manner known per se.
  • a saccharide preferably glucose
  • a preservative optionally 0.02-0.6 % by weight of a preservative
  • the reagent solution according to the invention may comprise as saccharide any mono- or disaccharide serving as energy source for the cells.
  • saccharide any mono- or disaccharide serving as energy source for the cells.
  • glucose is particularly preferred.
  • preservative preferably benzole acid and/or polyvinyl alcohol can be used.
  • the haemocytometrie examination of the cell suspension is performed as described in N. Eng. J. Med. 310, 1463 (1984). According to this method the cell suspension treated with the reagent solution is incubated, thereafter centrifuged, the supernatant is discarded, the sediment is admixed with a washing-diluting solution, and then staining is examined in a Btirker chamber.
  • the reagent solution according to the invention is presented preferably as a reagent kit.
  • the reagent kit comprises a reagent solution presented in dosage units (preferably filled into vials) and a washing-diluting solution presented in dosage units (preferably filled into vials), the volume ratio of said dosage units being 1 : (0.5-5).
  • any solution of known composition applied in cytologic examinations for this purpose can be used.
  • Hank's solution [J.H. Hank and R.E. Wallace: Proc. Soc. Exptl. Biol. Med. 71, 196 (1949)] proved to be particularly suitable.
  • the pH of the Hank's solution is adjusted to 7.2-7.4 (preferably to 7.3) with N-hydroxyethyl-piperazine-N'-2-ethanesulphonic acid.
  • the scope of the Invention also extends to the above reagent kit.
  • the major advantage of the invention is that the solutions applied in the examination are stable, those containing a preservative can be stored for at least 12 months without change, and can be presented in an immediately applicable form. Utilizing the reagent solution or reagent kit according to the invention diagnostic work can be performed more quickly and easily than before and the reliability and reproducibllity of the method is superior to those of the known ones.
  • Reagent kit is prepared from the following solutions: I. Reagent solution:
  • 0.1 g of polyvinyl alcohol is dissolved in 45 ml of hot distilled water, the solution is cooled to room temperature, and then 0.8 g of sodium chloride, 0.1 g of glucose, 0.02 g of benzole acid and 0.1 g of lysozyme are added. The pH of the resulting solution is adjusted to 6.9 with 1 molar Trie [tris(hydroaymethylamino-methane)] buffer solution, and then diluted to a final volume of 50 ml with distilled water.
  • washing-diluting solution 4 g of sodium chloride, 0.2 g of potassium chloride, 0.1 g of magnesium sulphate heptahydrate, 0.09 g of calcium chloride dihydrate, 0.03 g of disodium hydrophosphate dihydrate, 0.03 g of potassium dihydrophosphate and 0.5 g of glucose are dissolved in 350 ml of distilled water, and then 0.1 g of benzole acid is added to the solution.
  • the resulting solution is admixed with a solution of 0.175 g of sodium hydrocarbonate in 12.5 ml of distilled water.
  • the volume of the resulting solution is adjusted to 500 ml with distilled water, the solution is sterilized by passing through a G-5 sintered glass filter, and the filtrate is filled into vials each containing 5 ml of the solution.
  • Reagent kit is prepared from the following solutions: I. Reagent solution
  • the reagent solution has the same composition and is prepared in the same way as described in Example 1.
  • the volume of the resulting solution is adjusted to 500 ml with distilled water, thereafter the solution is sterilized by passing through a G-5 sintered glass filter, and the filtrate is filled into vials each containing 5 ml of the solution.
  • Example 3
  • Reagent kit is prepared from the following solutions: I. Reagent solution
  • the reagent solution has the same composition and is prepared in the same way as described in Example 1. II. Washing-diluting solution
  • the solution is admixed with a solution of 0.130 g of sodium hydrocarbonate in 10 ml of distilled water, and the pH of the resulting solution (Hank's solution) is adjusted to 7.3 with 5 ml of a 1 molar aqueous solution of N-hydroxyethyl-piperazine-N'-2-ethanesulphonic acid.
  • the volume of the resulting solution is adjusted to 500 ml with distilled water, the solution is sterilized by passing through a G-5 sintered glass filter, and the filtrate is filled into vials each containing 5 ml of the solution.
  • Reagent kit is prepared from the following solutions:
  • the reagent solution is prepared as described in Example 1 with the difference that 0.8 g of neutral red is applied.
  • the reagent solution is filtered, and the filtrate is filled into vials each containing 1 ml of the solution.
  • the washing-diluting solution has the same composition and is prepared in the same way as described in Example 1.
  • Example 5 Of the reagent kits described in Examples 1 to 4 one vial of the reagent solution and one vial of the washing-diluting solution are utilized for a single examination.
  • Example 5 Of the reagent kits described in Examples 1 to 4 one vial of the reagent solution and one vial of the washing-diluting solution are utilized for a single examination.

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Cell Biology (AREA)
  • Biochemistry (AREA)
  • Biotechnology (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Microbiology (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Ecology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

La présente invention se rapporte à une solution de réactif contenant du rouge neutre, qui sert à détecter des cellules ayant une activité phagocytaire. Ladite solution de réactif comprend 0,2 à 0,8 % en poids de rouge neutre, 0,1 à 0,25 % en poids de lysozyme, 0,8 à 2,0 % en poids de chlorure de sodium, 0,1 à 0,3 % en poids d'un saccharide et éventuellement 0,02 à 0,6 % en poids d'un agent conservateur dans une solution aqueuse ayant un pH compris entre 3,0 et 8,0, de préférence entre 3,5 et 5,0. La présente invention se rapporte également à un procédé permettant de détecter des cellules ayant une activité phagocytaire par l'utilisation d'une solution de réactif ayant la composition décrite ci-dessus.The present invention relates to a reagent solution containing neutral red, which is used to detect cells having phagocytic activity. Said reagent solution comprises 0.2 to 0.8% by weight of neutral red, 0.1 to 0.25% by weight of lysozyme, 0.8 to 2.0% by weight of sodium chloride, 0.1 to 0.3% by weight of a saccharide and optionally 0.02 to 0.6% by weight of a preservative in an aqueous solution having a pH of between 3.0 and 8.0, preferably between 3, 5 and 5.0. The present invention also relates to a method for detecting cells having phagocytic activity by the use of a reagent solution having the composition described above.

Description

REAGENT AUD METHOD FOR DETECTIHG CELLS WITH PHAGOCYTIC
ACTIVITY
The invention relates to a reagent and a method for detecting cells with phagocytic activity.
One of the major endeavours of medical science is to recognize developmental anomalies of foetl at the earliest possible stage of pregnancy.
It is well known that disorders of neural tube closure can be diagnostized by cell staining, since when such a disorder exists, cells with phagocytic activity appear in the amniotic fluid which can be detected by colour reaction. As staining agent neutral red is applied [N. Eng. J. Med. 310, 1463 (1984)]. Several problems have been emerged, however, when applying this method in the routine clinical diagnostics. It has been observed that the reliability of detection depends to a great extent on the quality of neutral red, i.e. the results obtained differ significantly when neutral red samples originating from different manufacturers are used. It is also difficult to prepare a solution of neutral red applicable for diagnostic purposes. The solution of neutral red in distilled water is of acidic character (the pH of a solution containing 1 mg/ml of neutral red is 3.1), and, owing to its acidity and hypoosmotic effect, this solution strongly damages the cells. None of the aqueous buffer and nutrient solutions utilized in routine clinical practice proved to be appropriate to dissolve neutral red, since after dissolution precipitation occurs. Organic solvents appeared to be less suitable; thus e.g. neutral red in ethanol solution does not give appropriate staining, whereas the dimethyl sulphoxide solution is difficult to sterilize.
The invention aims at providing a reagent solution and diagnostic method for detecting cells with phagocytic activity which is easy to apply and gives highly reliable results.
We have found that a neutral red-containing cytologic reagent solution should be approximately isoosmotic in relation to the potentially pathologic cells. It is also essential that the mixture of the reagent solu tion formed with the sample to be examined (amniotic fluid) should be almost neutral. It has also been observed that lysozyme enzyme significantly increases the stain uptake of the cells, moreover it Increases the stability of the reagent solution, too. Finally it has been found that the reagent solution should contain a saccharide as an energy source for the cells. When a reagent solution storable for a prolonged period of time is to be prepared, preferably a preservative should also be added to the mixture. Based on the above recognitions, the invention relates to a neutral red-containing reagent solution for detecting cells with phagocytic activity. The reagent solution according to the invention comprises 0.2-0.8 % by weight of neutral red, 0.1-0.25% by weight of lysozyme, 0.8-2.0 % by weight of sodium chloride, 0.1-0.3 % by weight of a saccharide and optionally 0.02-0.6 % by weight of a preservative in an aqueous solution with a pH of 3.0 to 8.0, preferably 3.5 to 5.0.
The invention also relates to a method for detecting cells with phagocytic activity by staining them with neutral red. According to the invention one proceeds as follows: 0.2-0.8 % by weight of neutral red, 0.1-0.25 % by weight of lysozyme, 0.8-2.0 % by weight of sodium chloride, 0.1-0.3 % by weight of a saccharide (preferably glucose) and optionally 0.02-0.6 % by weight of a preservative are dissolved in water, the pH of the solution is adjusted to 3.0-8.0, preferably 3.5-5.0, the solution is sterilized by filtration, thereafter one part by volume of the resulting reagent solution is admixed with 3-8 parts by volume of a sample to be examined, preferably amniotic fluid, and the resulting cell suspension is examined in a haemocytometer in a manner known per se.
The reagent solution according to the invention may comprise as saccharide any mono- or disaccharide serving as energy source for the cells. Of the saccharides glucose is particularly preferred. As preservative preferably benzole acid and/or polyvinyl alcohol can be used.
The haemocytometrie examination of the cell suspension is performed as described in N. Eng. J. Med. 310, 1463 (1984). According to this method the cell suspension treated with the reagent solution is incubated, thereafter centrifuged, the supernatant is discarded, the sediment is admixed with a washing-diluting solution, and then staining is examined in a Btirker chamber. For clinical examinations the reagent solution according to the invention is presented preferably as a reagent kit. The reagent kit comprises a reagent solution presented in dosage units (preferably filled into vials) and a washing-diluting solution presented in dosage units (preferably filled into vials), the volume ratio of said dosage units being 1 : (0.5-5).
As washing-diluting solution any solution of known composition applied in cytologic examinations for this purpose can be used. Hank's solution [J.H. Hank and R.E. Wallace: Proc. Soc. Exptl. Biol. Med. 71, 196 (1949)] proved to be particularly suitable. According to a particularly preferred method the pH of the Hank's solution is adjusted to 7.2-7.4 (preferably to 7.3) with N-hydroxyethyl-piperazine-N'-2-ethanesulphonic acid. The scope of the Invention also extends to the above reagent kit.
The major advantage of the invention is that the solutions applied in the examination are stable, those containing a preservative can be stored for at least 12 months without change, and can be presented in an immediately applicable form. Utilizing the reagent solution or reagent kit according to the invention diagnostic work can be performed more quickly and easily than before and the reliability and reproducibllity of the method is superior to those of the known ones.
The invention is elucidated in detail by the aid of the following non-limiting Examples. Example 1
Reagent kit is prepared from the following solutions: I. Reagent solution:
0.1 g of polyvinyl alcohol is dissolved in 45 ml of hot distilled water, the solution is cooled to room temperature, and then 0.8 g of sodium chloride, 0.1 g of glucose, 0.02 g of benzole acid and 0.1 g of lysozyme are added. The pH of the resulting solution is adjusted to 6.9 with 1 molar Trie [tris(hydroaymethylamino-methane)] buffer solution, and then diluted to a final volume of 50 ml with distilled water. A solution of 0.2 g of neutral red (3-amino-7-dimethylamino-2-methyl-phenozine hydrochloride) in 50 ml of distilled water is added to the above solution, the mixture is well stirred, sterilized by passing through a G-5 sintered glass filter, and the filtrate is filled into vials each containing 1 ml of the solution. II. Washing-diluting solution 4 g of sodium chloride, 0.2 g of potassium chloride, 0.1 g of magnesium sulphate heptahydrate, 0.09 g of calcium chloride dihydrate, 0.03 g of disodium hydrophosphate dihydrate, 0.03 g of potassium dihydrophosphate and 0.5 g of glucose are dissolved in 350 ml of distilled water, and then 0.1 g of benzole acid is added to the solution. The resulting solution is admixed with a solution of 0.175 g of sodium hydrocarbonate in 12.5 ml of distilled water. The mixture is well stirred, then the pH of the resulting solution (Hank's solution) is adjusted to 7.3 with 5 ml of a 1 molar aqueous solution of N-hydroxyethyl-piperazine-N'-2-ethanesulphonic acid.
The volume of the resulting solution is adjusted to 500 ml with distilled water, the solution is sterilized by passing through a G-5 sintered glass filter, and the filtrate is filled into vials each containing 5 ml of the solution.
Example 2
Reagent kit is prepared from the following solutions: I. Reagent solution
The reagent solution has the same composition and is prepared in the same way as described in Example 1.
II. Washing-diluting solution
4 g of sodium chloride, 0.2 g of potassium chloride, 0.1 g of magnesium sulphate heptahydrate, 0.03 g of disodium hydrophosphate, 0.03 g of potassium dihydrophosphate, 0.5 g of glucose and 0.09 g of calcium chloride dihydrate are dissolved in 350 ml of distilled water, and 0.5 g of benzolc acid is added to the solution. The resulting solution is admixed with a solution of 2.4 g of sodium hydrocarbonate in 30 ml of distilled water. The mixture is well stirred, then the pH of the resulting solution (Hank's solution) is adjusted to 7.3 with 5 ml of a 1 molar aqueous solution or N-hydroxyethyl-piperazine-N'-2-ethanesulphonic acid.
The volume of the resulting solution is adjusted to 500 ml with distilled water, thereafter the solution is sterilized by passing through a G-5 sintered glass filter, and the filtrate is filled into vials each containing 5 ml of the solution. Example 3
Reagent kit is prepared from the following solutions: I. Reagent solution
The reagent solution has the same composition and is prepared in the same way as described in Example 1. II. Washing-diluting solution
4 g of sodium chloride, 0.2 g of potassium chloride, 0.1 g of magnesium sulphate heptahydrate, 0.03 g of disodium hydrophosphate dihydrate, 0.03 g of potassium dihydrophosphate, 0.5 g of glucose and 0.09 g of calcium chloride dihydrate are dissolved in 350 ml of distilled water, thereafter 0.6 g of sodium benzoate is added to the solution. The solution is admixed with a solution of 0.130 g of sodium hydrocarbonate in 10 ml of distilled water, and the pH of the resulting solution (Hank's solution) is adjusted to 7.3 with 5 ml of a 1 molar aqueous solution of N-hydroxyethyl-piperazine-N'-2-ethanesulphonic acid. The volume of the resulting solution is adjusted to 500 ml with distilled water, the solution is sterilized by passing through a G-5 sintered glass filter, and the filtrate is filled into vials each containing 5 ml of the solution. Example 4
Reagent kit is prepared from the following solutions:
I. Reagent solution
The reagent solution is prepared as described in Example 1 with the difference that 0.8 g of neutral red is applied. The reagent solution is filtered, and the filtrate is filled into vials each containing 1 ml of the solution.
II. Washing-diluting solution
The washing-diluting solution has the same composition and is prepared in the same way as described in Example 1.
Of the reagent kits described in Examples 1 to 4 one vial of the reagent solution and one vial of the washing-diluting solution are utilized for a single examination. Example 5
5 ml of amniotic fluid are admixed with one vial (1 ml) of the reagent solution described in Example 1. The mixture is incubated for 15 minutes, then centrifuged, the supernatant is discarded, and the sediment is admixed with 0.5 to 2.0 ml of the washing-diluting solution described in Example 1. The stained sample is examined in a Bürker chamber.

Claims

What we claim is:
1. A neutral red-containing reagent solution for detecting cells with phagocytic activity, which comprises 0.2-0.8 % by weight of neutral red, 0.1-0.25 % by weight of lysozyme, 0.8-2.0 % by weight of sodium chloride, 0.1-0.3 % by weight of a saccharide and optionally 0.02-0.6 by weight of a preservative in an aqueous solution with a pH of 3.0 to 8.0, preferably 3.5 to 5.0.
2. A reagent solution as claimed in claim 1, in which the saccharide is a mono- or disaccharide serving as energy source for the cells, preferably glucose.
3. A reagent solution as claimed in claim 1 or 2, in which the preservative is benzole acid and/or polyvinyl alcohol.
4. A method for detecting cells with phagocytic activity by staining them with neutral red , characterized in that 0.2-0.8 % by weight of neutral red, 0.1-0.25 % by weight of lysozyme, 0.8-2.0 % by weight of sodium chloride, 0.1-0.3 % by weight of a saccharide, preferably glucose, and optionally 0.02-0.6 % by weight of a preservative are dissolved in water, the pH of the solution is adjusted to 3.0-8.0, preferably 3.5-5.0, the solution is sterilized by filtration, thereafter one part by volume of the resulting reagent solution is admixed with 3-8 parts by volume of a sample to be examined, preferably amniotic fluid, and the resulting cell suspension is examined in a haemocytometer in a manner known per se.
5. Reagent kit for the detection of cells with phagocytic activity, which comprises a reagent solution as claimed in any of claims 1 to 3 presented in dosage units and a washing-diluting solution presented in dosage units, and the volume ratio of said dosage units is
1 : (0.5-5).
6. A reagent kit as claimed in claim 5, which comprises vials containing 1 ml each of the reagent solution and vials containing 5 ml each of the washing-diluting solution.
7. A reagent kit as claimed in claim 5 or 6, which comprises as washing-diluting solution a Hank's solution the pH of which is adjusted to 7.2-7.4 with N-hydroxyethyl-piperazine-N'-2-ethanesulphonic acid.
EP19880902576 1987-03-27 1988-03-22 Reagent and method for detecting cells with phagocytic activity Withdrawn EP0345295A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
HU134087 1987-03-27
HU134087A HU197450B (en) 1987-03-27 1987-03-27 Reagent and method for detecting cells of phagocyte activity

Publications (1)

Publication Number Publication Date
EP0345295A1 true EP0345295A1 (en) 1989-12-13

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EP19880902576 Withdrawn EP0345295A1 (en) 1987-03-27 1988-03-22 Reagent and method for detecting cells with phagocytic activity

Country Status (5)

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EP (1) EP0345295A1 (en)
JP (1) JPH03500961A (en)
AU (1) AU1488588A (en)
HU (1) HU197450B (en)
WO (1) WO1988007583A1 (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105651579A (en) * 2015-12-07 2016-06-08 上海太阳生物技术有限公司 Alpha-naphthol butyrate esterase staining kit

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE2240672C2 (en) * 1972-08-18 1974-08-22 Boehringer Mannheim Gmbh, 6800 Mannheim Method and device for staining biological material

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO8807583A1 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105651579A (en) * 2015-12-07 2016-06-08 上海太阳生物技术有限公司 Alpha-naphthol butyrate esterase staining kit
CN105651579B (en) * 2015-12-07 2018-10-23 上海太阳生物技术有限公司 A kind of iophenoxic acid naphthol ester enzyme staining reagent kit

Also Published As

Publication number Publication date
HU197450B (en) 1989-03-28
HUT46145A (en) 1988-09-28
WO1988007583A1 (en) 1988-10-06
AU1488588A (en) 1988-11-02
JPH03500961A (en) 1991-03-07

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Inventor name: PAPP, ZOLTAN

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