Spectral differentiation of histological stains
A technology for histology and stains, applied in the field of spectral differentiation of histological stains, can solve the problems of difficult to obtain complex mixtures, laborious, time-consuming and resource-consuming
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example 1
[0187] Example 1: Stain sample preparation
[0188] Romanowski stains, including Wright-Giemsa (WG) stains and Meglinger (MG) stains, are prepared to include stain compositions within specifications and intentionally within outside the specification. Specifically, the prepared stain was below specification, within specification but below target, on target, within specification but above target, and above specification. In Table 1 ( figure 1 ) provides peak absorbance measurements at approximately 655 nm for prepared 1:200 dilutions of on-target and off-target stains in deionized water (DI).
example 2
[0189] Example 2: Determination of the lower limit of absorbance
[0190] determined at 655nm and 517nm respectively and at figure 2 and image 3 The initial absorbance of different preparations of WG stain and MG stain is provided in .
[0191] Table 2
[0192] WG sample 655nm 517nm 50% lower spec 0.481 0.330 67% low spec 0.656 0.490 83% low spec 0.798 0.620 100% low spec 0.951* 0.765* Target 1.052 0.880 100% high specification 1.128 0.978 110% high specification 1.249 1.104
[0193] table 3
[0194] MG sample 655nm 517nm 50% lower spec 0.298 0.257 67% low spec 0.398 0.362 83% low spec 0.499 0.481 100% low spec 0.600 0.597 Target 0.682 0.713 100% high specification 0.779 0.840 110% high specification 0.888 0.997
[0195]Absorbance measurements of the stains were performed according to the following procedure using a 1:200 dilution of each ...
example 3
[0205] For comparison of cell staining between different WG-stained samples with different absorbance values and different dye concentrations, exemplary images are provided ( figure 2 ). Exemplary images are also provided for comparison of cell staining between samples of different MG stains with different absorbance values and different dye concentrations ( image 3 ). Example 3: Spectral Resolution of Different Romanowski Stains
[0206] Use spectral absorbance measurements to resolve Wright-Giemsa stains from Meglingham stains. Absorption spectra were measured for the stains in the spectral range from 200 nm to 800 nm with a step size of 1.0 nm. The exact position of the peak in the spectral range of 650nm-665nm was identified with an accuracy of ±1.0nm. For the particular staining preparation and absorbance measurement used in this example, the stain was identified as Wright-Giemsa when the absorbance peak position was at λ = 659 nm ± 1 nm, whereas when the peak p...
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