A New Multiparameter Separator for Microscopic Particles and Biological Cells

Steinkamp, John A.; Fulwyler, Mack J.; Coulter, J. R.; Hiebert, R. D.; Horney, J. L.; Mullaney, P. F.

doi:10.1063/1.1686375

Cited by 208 publications

(67 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Flow microfluorometry.-A flow microfluorometer with a laser wave-length setting of 457 ,um was used to measure the DNA content of individual cells in suspension (Steinkamp et al, 1973). Cells were stained with mithramycin (Grdina et al, 1978a).…”

Section: Methodsmentioning

confidence: 99%

Phase-specific cytotoxicity in vivo of hydroxyurea on murine fibrosarcoma cells synchronized by centrifugal elutriation

Grdina¹,

Sigdestad²,

Peters³

1979

Br J Cancer

View full text Add to dashboard Cite

show abstract

Section: Methodsmentioning

confidence: 99%

Phase-specific cytotoxicity in vivo of hydroxyurea on murine fibrosarcoma cells synchronized by centrifugal elutriation

Grdina¹,

Sigdestad²,

Peters³

1979

Br J Cancer

View full text Add to dashboard Cite

show abstract

“…Flow cytometry is the only technology in which rapid single cell multiparameter analysis and sorting can be accomplished (2)(3)(4)(5)(6)(7)(8). In keeping with Mack's great contributions to flow cytometry, we present an extension of its capabilities into the infrared (IR).…”

mentioning

confidence: 99%

Flow cytometer in the infrared: Inexpensive modifications to a commercial instrument

et al. 2005

View full text Add to dashboard Cite

Background: The application of molecules that fluoresce in the infrared (IR) region to measure cell products would be enhanced by a flow cytometer capable of measuring them. To our knowledge, none exist at this time. Accordingly, we have developed such an instrument. Methods: A Becton Dickinson LSR flow cytometer was modified to include a small 785-nm IR diode laser the size of a C cell battery with 44-mW output power. The instrument was modified further to accommodate this laser in addition to a 405-nm solid-state laser, a 488-nm air-cooled argon laser, and a 658-nm solid-state laser. Because the IR laser is dangerous to the eye, the laser beams were viewed for optical alignment using a CCD camera and video monitor. An avalanche photodiode was used in place of a photomultiplier tube because its detection sensitivity in the IR region is superior. Results: To assess performance, scatter and florescence measurements were made using microspheres that fluoresce in the IR region, and human leukocytes were stained

show abstract

“…The first lens collected the emitted fluorescence and focused it at infinity through a filter to the second lens, which refocused the light onto a PMT pinhole aperture. Projector lenses also have been used for quantifying multicolor fluorescence (28,301. To improve the light-collecting efficiency, a plano-convex lens can be attached to the flow chamber to cancel the divergence of light rays caused by refraction at the glass/ air interface (6).…”

mentioning

confidence: 99%

A modular detector for flow cytometric multicolor fluorescence measurements

1987

View full text Add to dashboard Cite

A modular detector for measuring multicolor fluorescence from cells illuminated by single or multiple lasers has been developed for flow cytometers. Motion picture projector, camera, and CCTVlvideo lenses were evaluated for use in the detector by comparing their physical characteristics, image quality, and light collection efficiencies. A 25-mm focal length F/0.95 CCTV lens was selected, based on our critera and test results. The detector was constructed out of square aluminum extrusion channels. A CCTV lens mounted on the outside of the first channel collected light emitted from cells and collimated it towards filters and secondary CCTV lenses located in each channel. The secondary lenses functioned as relay optics for directing and focusSeveral optical systems have been designed for measuring light scatter and fluorescence in flow cytometers (FCM). Early methods employed bright-fieladark-field microscopic on-axis measurements for quantifying absorption, light scatter (12), and fluorescence (11). In these detection systems, high numerical aperture (NA) objective lenses were used to magnify and image the cell stream onto apertures in front of photomultiplier tubes (PMTs). Dichroic color-separating filters were used to divide the absorption, light scatter, and fluorescence. A similar dark-field illumination instrument for quantifying fluorescence and light scatter was constructed around a 10 x low-power microscope (27).Epiilluminators (15) have been used to measure fluorescence (5,241 and light scatter (23) and they have been used with flow chambers that have electronic particlevolume sensing (9) and cell-sorting (13) capability. High-NA objectives serve as both the excitation and emission collection lens. The illuminating light is reflected by a n excitation dichroic mirror and imaged onto the cell stream. The longer-wavelength fluorescence emission is then collected by the same lens and transmitted by the dichroic mirror for imaging onto PMT detectors. Multibeam, sequential excitation also has been used to illuminate cells for measuring absorption, scatter, and ing light onto pinhole spatial filters for measurement by photomultipliers. The detector design allowed any number of channels to be connected together and the versatility for making simultaneous or sequential measurements. Measurements on lymphocytes labeled with four monoclonal antibodies conjugated to fluorescent dyes and measurements on viable tumor cells stained for DNA content and with three fluorescent-labeled antibodies were used to demonstrate the detector's capabilities.Key terms: Fluorescence detector, immunofluorescence, lymphocytes, tumor-cell analysis multicolor fluorescence (3). Recently, the fluorescence emission spectra of cells have been recorded using a monochromator connected to a vertical-illuminator flow cytometer (25).The orthogonal dark-field method uses a lens placed perpendicular to the laser-beadcell-stream axes to collect the emitted fluorescence and light scatter. Two projector lenses were originally used as relay optics (1:l...

show abstract

A New Multiparameter Separator for Microscopic Particles and Biological Cells

Cited by 208 publications

References 17 publications

Phase-specific cytotoxicity in vivo of hydroxyurea on murine fibrosarcoma cells synchronized by centrifugal elutriation

Phase-specific cytotoxicity in vivo of hydroxyurea on murine fibrosarcoma cells synchronized by centrifugal elutriation

Flow cytometer in the infrared: Inexpensive modifications to a commercial instrument

A modular detector for flow cytometric multicolor fluorescence measurements

Contact Info

Product

Resources

About