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Biomedical subjects

L D Kamentsky

Publications and source records attributed to L D Kamentsky.

4 recordsLinked to original sources
PubMed1997

Methods for automatic multiparameter analysis of fluorescence in situ hybridized specimens with a laser scanning cytometer.

Multiparameter laser scanning cytometry has been applied to the automatic counting of probe spots and the simultaneous measurement of cellular DNA for fluorescence in situ hybridization (FISH) prepared specimens counterstained with propidium iodide. Relatively low resolution imaging, highly variable probe fluorescence, spectral overlap of probe with counterstain fluorescence, and autofluorescence required the development of an image processing method to detect and isolate FISH probe spots. Inability to properly apportion detected probe spots because of overlapping probe spot images in the same cell required development of a method to eliminate cell data whenever spots in that cell could not be reliably isolated. Laser scanning cytometry incorporating these methods to determine per cell probe spot count and DNA is demonstrated on tissue cultures and peripheral blood cells using different centromeric FISH probes with either FITC or Spectrum Green labeling.

Cells, Cultured↗
PubMed1991

Microscope-based multiparameter laser scanning cytometer yielding data comparable to flow cytometry data.

We describe a computer-controlled 10 microns spot size laser scanning cytometer for making multiple wavelength fluorescence and scatter measurements of unconstrained cells on a surface such as a microscope slide. Designated areas of slides placed on a microscope stage are automatically scanned, and cells which generate above-threshold scatter or fluorescence values are found and individually processed to determine a list of measurement parameters. For each fluorescence or scatter measurement parameter, this list contains the integrated and peak values and bit pattern images of a scan window centered on the cell. The measurement time, the position of the cell on the slide, and two segmentation indices are also included in the list. Measurement time, cell position, and properties derived from the bit patterns are used interchangeably with integrated or peak measurement values as coordinates of multiproperty displays. Cells may be selected for counting, data display in various forms, or visual observation based on their meeting complex criteria among a chain of two property screens. Cells with selected properties may be viewed during an experiment or retrospectively. A designated specimen field may be repeatedly remeasured to perform kinetic cell studies. An argon ion and a HeNe- based laser instrument have been constructed and software has been written and evaluated with the specific goal of increasing the precision of propidium iodide-stained cellular DNA measurements. Some of the capabilities of the instrument and its current performance are described.

Algorithms↗
PubMedDate not supplied

Slide-based laser scanning cytometry.

OBJECTIVE: To show that laser scanning cytometry (LSCM) can provide data equivalent to flow cytometry (FCM) data and furnish a number of benefits, including cell relocation for visualization and several additional measurement features that may make it more suitable than FCM for pathology laboratories. STUDY DESIGN: A laser scanning cytometer, the LSC, was developed. Several instruments, at sites in the United States and Japan during the last two years, provided data characterizing the instrument and its usefulness. RESULTS: Data describing the sensitivity, precision, accuracy, utility of added measurement features and cell relocation capabilities of the LSC are presented. The data illustrate the applicability of the LSC to multiparameter DNA ploidy studies, resolution of phases of the cell cycle and cytogenetics. CONCLUSION: Because it is microscope based and measures cells on a slide, not in a flow chamber; records the position of each cell on the slide; and has higher resolution, LSCM provides a number of benefits that may make it more suitable than FCM for pathology laboratories.

Animals↗