Method and apparatus for identifying reticulocytes within a blood sample
A method and apparatus for identifying reticulocytes within a blood sample is provided. The method includes the steps of: a) depositing the sample into an analysis chamber adapted to quiescently hold the sample for analysis, and the chamber has a known or determinable height extending between the interior surfaces of panels, which height is such that at least one red blood cell, or an aggregate of red blood cells, within the sample contacts both of the interior surfaces; b) admixing a supravital dye with the sample, which dye is operable to cause reticulin to fluoresce when excited by light of one or more predetermined wavelengths; c) imaging the sample using light that includes the one or more predetermined wavelengths that cause reticulin to fluoresce; d) imaging the sample using light that is absorbed by hemoglobin to produce values of optical density on a per image unit basis; and e) identifying reticulocytes within the sample using the image of the sample created with light that causes the dyed reticulin to fluoresce, and using the per image unit optical density values.
1. A method for identifying reticulocytes within a blood sample, comprising the steps of:
depositing the sample into an analysis chamber adapted to quiescently hold the sample for analysis;
admixing a supravital dye with the sample, which dye is operable to cause reticulin within reticulocytes within the sample to fluoresce when excited by light of one or more predetermined wavelengths;
imaging the sample using light that includes the one or more predetermined wavelengths that cause reticulocytes within the sample to fluoresce;
imaging the sample using light at one or more predetermined wavelengths that is absorbed by hemoglobin to produce values of optical density on a per image unit basis; and
identifying reticulocytes within the sample using the image of the sample created with light at the one or more predeteimined wavelengths that causes the dyed reticulin to fluoresce.
2. An apparatus for identifying reticulocytes within a blood sample, comprising:
an analysis chamber adapted to quiescently hold the sample for analysis;
an imaging unit that includes an illuminator and an image dissector, which unit is operable to image one or more red blood cells along predetermined wavelengths of light, and produce image signals representative of such imaged red blood cells, wherein some of the predetermined wavelengths are such that reticulocytes within the sample fluoresce when the sample mixed with a supravital dye is illuminated by the wavelengths, and wherein other of the wavelengths are such that they are absorbed by hemoglobin to produce values of optical density; and
a programmable analyzer in communication with the imaging unit, which analyzer is adapted to identify reticulocytes, using the image of the sample created with light at the predetermined wavelengths.
3. A method for determining red blood cell hemoglobin concentration within a blood sample, comprising the steps of:
depositing the sample into an analysis chamber adapted to quiescently hold the sample for analysis, the chamber having a known or determinable height;
imaging at least one red blood cell;
determining an optical density of at least a portion of the imaged red blood cell; and
determining the hemoglobin concentration of one or more red blood cells within the sample, using the determined optical density.
4. A method for determining a cell volume of a red blood cell within a blood sample, comprising the steps of:
depositing the sample into an analysis chamber adapted to quiescently hold the sample for analysis, the chamber having a known or determinable height;
imaging at least one red blood cell in the sample, and producing image signals;
determining an optical density of a portion of the at least one red blood cell; and
determining a volume of the red blood cell using the determined optical density, and the height of the chamber.