Automated systems and methods for preparing biological specimens for examination

A method of preparing a biological specimen on a substrate for examination, the method comprising:

• (a) positioning the substrate with respect to a surface so that the biological specimen faces the surface, and so that the substrate and the surface are substantially parallel and form a separation of at least about 100 microns;
• (b) sequentially dispensing (i) a first fixative solution, (ii) a first stain solution, (iii) a second stain solution, and (iv) a first rinse solution into the separation between the substrate and the surface in an amount sufficient to contact the specimen and the surface;
• and
• (c) after dispensing each one of solutions (i), (ii), (iii), and (iv) in step (b), and before dispensing the next one of solutions (i), (ii), (iii), and (iv) in step (b):
• performing at least a first agitation cycle, wherein the first agitation cycle comprises changing a distance between the substrate and surface while the dispensed solution contacts the specimen for the duration of the first agitation cycle;
• and
• removing the dispensed solution from the separation and from contacting the specimen.

A method of preparing a biological specimen on a substrate for examination, the method comprising:

• (a) positioning the substrate with respect to a surface so that the specimen faces the surface, and so that the substrate is positioned to form a separation between the surface and at least a portion of the substrate of at least about 100 microns;
• (b) performing a fixing phase comprising:
• (i) dispensing a fixative into the separation between the substrate and the surface in an amount sufficient to contact the specimen and the surface;
• (ii) performing at least a first agitation phase, wherein the first agitation phase comprises changing a distance between the substrate and surface while the fixative is contacting the specimen for the duration of the first agitation phase;
• and
• (iii) removing the fixative from the separation and the specimen;
• (c) performing a first staining phase comprising:
• (i) dispensing a first stain into the separation between the substrate and the surface in an amount sufficient to contact the specimen and the surface;
• (ii) performing at least a second agitation phase, wherein the second agitation phase comprises changing the distance between the substrate and surface while the first stain is contacting the specimen for the duration of the second agitation phase;
• and
• (iii) removing the first stain from the separation and the specimen;
• (d) performing a second staining phase comprising:
• (i) dispensing a second stain into the separation between the substrate and the surface in an amount sufficient to contact the specimen and the surface;
• (ii) performing at least a third agitation phase, wherein the third agitation phase comprises changing the distance between the substrate and surface while the second stain is contacting the specimen for the duration of the third agitation phase;
• and
• (iii) removing the second stain from the separation and the specimen;
• and
• (e) performing a first rinse phase comprising:
• (i) dispensing a first rinse into the separation between the substrate and the surface in an amount sufficient to contact the specimen and the surface;
• (ii) performing at least a fourth agitation phase, wherein the fourth agitation phase comprises changing the distance between the substrate and surface while the first rinse is contacting the specimen for the duration of the fourth agitation phase;
• and
• (iii) removing the first rinse from the separation and the specimen.