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Hematoxylin solution comprising chloride and sulphate, and methods of preparation and use

A kind of technology of sulfate ion and hematoxylin, applied in the field of hematoxylin preparation

Pending Publication Date: 2017-08-29
VENTANA MEDICAL SYST INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, hematoxylin autostaining solutions have unique requirements that cannot be met by formulations suitable for manual staining

Method used

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  • Hematoxylin solution comprising chloride and sulphate, and methods of preparation and use
  • Hematoxylin solution comprising chloride and sulphate, and methods of preparation and use
  • Hematoxylin solution comprising chloride and sulphate, and methods of preparation and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] ANS hematoxylin is based on the use of Al 2 (SO 4 ) 3 ·nH 2 O acts as a mordant and has been specifically optimized for automated H&E staining using a VentanaSYMPHONY autostainer. The formulations are based on those disclosed in US Patent 8,551,731. The hematoxylin preparation is prepared according to the following formulation:

[0062]

[0063] The composition is prepared according to the following procedure:

[0064] 1. Add / dissolve hematoxylin into a solution of water and glycol.

[0065] 2. Add / dissolve sodium iodate.

[0066] 3. Add / dissolve Al 2 (SO 4 ) 3 ·16H 2 o

[0067] 4. Add / dissolve hydroquinone and β-cyclodextrin

[0068] This composition is used as a benchmark for automated dyeing performance as it is the best known composition. The composition is capable of delivering proper staining on slides in about two minutes. While the composition stained well, with the correct shade and depth, and had sufficient long-term stability; it was not idea...

Embodiment 2

[0071] Prepare Cl with 1 / 0 - / SO 4 2- Molar ratio (i.e. 100% Cl - ) hematoxylin preparation. In addition to using AlCl 3 Hexahydrate instead of Al 2 (SO 4 )3 16H 2 Except for O, the same components and concentrations as in Example 1 were used.

[0072]

[0073] The composition was observed to be very stable, but also had very slow kinetics during dyeing. Therefore, the staining is very light under normal operating conditions.

Embodiment 3

[0075] Prepare Cl with 1 / 1 - / SO 4 2- Molar ratio (ie 50% Cl - ) hematoxylin preparation. In such formulations, including SO 4 2- , because it is completely removed from ANS-Cl, resulting in poor staining quality (depth and shade of staining) and unacceptable kinetics (eg too slow for automated staining). In addition, the concentration of hematoxylin was also increased to increase the kinetics, which is thought to be influenced by Cl - suppressed by the existence of

[0076]

[0077] The formulations showed high quality dyeing and adequate dyeing rates. After several experiments, it was concluded that higher concentrations of hematoxylin compared to ANS could be used to ensure staining kinetics matched the ANS benchmark. The compositions were observed to be sufficiently stable over extended periods of time without including β-cyclodextrin or hydroquinone included in the ANS. It was also observed that this formulation had a significantly delayed onset of precipitati...

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PUM

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Abstract

This disclosure relates to hematoxylin staining formulations and particularly to formulations for use in autostainers. The disclosed compositions were discovered to possess atypical stability under storage while having high stain quality and sufficiently fast staining performance. The disclosed hematoxylin staining compositions include a solvent system, hematoxylin, a chemical oxidant, and a mordant. Illustrative embodiments also have a Cl- / SO4- molar ratio of between about 2.5 / 1 and about 1 / 4.

Description

technical field [0001] The present invention relates to compositions and methods for histochemical staining of biological samples. More specifically, the present invention relates to hematoxylin formulations with enhanced degradation stability. Background technique [0002] Hematoxylin has been described as the most important and commonly used dye in histology, histochemistry, histopathology and cytology. Several histochemical staining protocols, including hematoxylin and eosin (H&E) staining and Papanicolaou (PAP) staining) rely on the dye hematoxylin to stain cytological and tissue samples. Specifically, pathologists use hematoxylin staining of cell nuclei to detect the presence of malignant and / or metastatic cells in tumor biopsy samples. Hematoxylin, also known as haematoxylin, natural black 1, or C.I. 75290m, is a naturally occurring compound found in the red heartwood of trees of the genus Hematoxylon, such as the loggerhead tree . Hematoxylin is not an active ingr...

Claims

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Application Information

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IPC IPC(8): G01N1/30
CPCG01N1/30G01N21/253G01N1/312G01N2001/302
Inventor H·巴奈特E·达兰特E·L·克莱因M·林奇K·普理查德
Owner VENTANA MEDICAL SYST INC
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