Live-cell assay for detection of apoptosis by dual-laser flow cytometry using Hoechst 33342 and 7-amino-actinomycin D

2007 ◽  
Vol 2 (1) ◽  
pp. 187-190 ◽  
Author(s):  
Ingrid Schmid ◽  
Christel Uittenbogaart ◽  
Beth D Jamieson
Keyword(s):  
Flow Cytometry ◽  
Actinomycin D ◽  
Live Cell ◽  
Hoechst 33342 ◽  
Cell Assay ◽  
Dual Laser

scholarly journals hFUT1-Based Live-Cell Assay To Profile α1-2-Fucoside-Enhanced Influenza Virus A Infection

2020 ◽  
Vol 15 (4) ◽  
pp. 819-823 ◽  
Author(s):  
Senlian Hong ◽  
Geramie Grande ◽  
Chenhua Yu ◽  
Digantkumar G. Chapla ◽  
Natalie Reigh ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Live Cell ◽  
Influenza Virus A ◽  
Cell Assay

scholarly journals Detection and analysis by dual-laser flow cytometry of bacteriophage T4 DNA insideEscherichia coli

Cytometry ◽  
1991 ◽  
Vol 12 (2) ◽  
pp. 167-171 ◽  
Author(s):  
Cynthia A. Sanders ◽  
David M. Yajko ◽  
Patricia S. Nassos ◽  
William C. Hyun ◽  
Mack J. Fulwyler ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Bacteriophage T4 ◽  
Dual Laser

461 Prostate stem cell isolation using Hoechst 33342 flow cytometry

2004 ◽  
Vol 3 (2) ◽  
pp. 118
Author(s):  
P. Gilmore ◽  
R. Bhatt ◽  
C. Hart ◽  
V. Ramani ◽  
N. George ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Stem Cell ◽  
Cell Isolation ◽  
Hoechst 33342 ◽  
Stem Cell Isolation

scholarly journals Intermittent blood flow in the KHT sarcoma--flow cytometry studies using Hoechst 33342

1990 ◽  
Vol 62 (2) ◽  
pp. 195-200 ◽  
Author(s):  
AI Minchinton ◽  
RE Durand ◽  
DJ Chaplin
Keyword(s):  
Flow Cytometry ◽  
Blood Flow ◽  
Hoechst 33342

110 COMPARISON OF ANDROMED®, BIOXCELL®, AND BOTU-BOV® EXTENDERS FOR CRYOPRESERVATION OF BULL SEXED SEMEN

2007 ◽  
Vol 19 (1) ◽  
pp. 172 ◽  
Author(s):  
M. Q. Braga ◽  
R. V. R. Franco ◽  
L. F. Rodrigues ◽  
G. Galeli ◽  
K. M. Oliveira ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Semen Quality ◽  
Membrane Damage ◽  
Optical Microscope ◽  
Hoechst 33342 ◽  
Tukey Test ◽  
Progressive Motility ◽  
Significant Difference ◽  
Sexed Semen ◽  
Fort Collins

Sexing semen has become a worldwide technology now available in many countries through the use of flow cytometry for sexing mammal sperms (Johnson and Welch 1999 Theriogenology 52, 1323–1341). Because straws containing sexed semen have a low concentration, any condition that either improves or decreases freezing capabilities will considerably change semen quality. During cryopreservation, spermatozoa have been described as undergoing many changes that lead to membrane damage, which may result in decreased fertility (Watson 2000 Reprod. Fertil. Dev. 6 (Suppl 1), 481). Since many cryoprotectants are available on the market, the objective of the present study was to compare 3 different extenders for freezing sex-sorted semen. For this study, 25 ejaculates were collected from 8 bulls of different breeds, diluted, then dyed with Hoechst 33342 (Schenk et al. 1999 Theriogenology 52, 1375–1391), and sexed by flow cytometry (SX MoFlo®; DaKoCytomation, Inc., Fort Collins, CO, USA). After being cooled at 4°C for 1 h and 30 min, the sexed semen was centrifuged and diluted in AndroMed® (Minitüb, Tiefenbach, Germany), Bioxcell® (IMV, Aigle, France), or Botu-Bov® (Biotech Botucatu, Ltda., Sao Paulo, Brazil); the semen was packaged at 3 million total sperm in 0.25-mL straws and frozen in an automatic freezer (Digit cool 5300® IMV). To evaluate the freezing quality, the straws were thawed and incubated at 35°C for 15 min. The progressive motility was observed through an optical microscope (Coleman 200T). The statistical analyses were done using the SAS program (SAS Institute, Inc., Cary, NC, USA) and the Tukey test (P ≤ 0.05). Results show that there was no statistical difference between Bioxcell and AndroMed extenders (P ≤ 0.05). However, Botu-Bov extender showed a significant difference when compared with Bioxcell and AndroMed (see Table 1). It is also important to point out that 40% of the samples frozen with AndroMed showed non-aligned movement. Even though few ejaculates were used for this study, preliminary results showed that Bioxcell seemed to be the most suitable for freezing bull sexed semen. Table 1. Percentage of progressively motile spermatozoa after thawing

Using dual-laser flow cytometry for monitoring phytoplankton composition and integrity

1995 ◽  
Author(s):  
Harald Schaefer ◽  
Wolfgang Beisker ◽  
Christian Steinberg
Keyword(s):  
Flow Cytometry ◽  
Phytoplankton Composition ◽  
Dual Laser

Use of Pacific Orange™ Dye with Qdot® Nanocrystals and Other Violet-Excited Dyes in Polychromatic Flow Cytometry.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 3901-3901
Author(s):  
Gayle M. Buller ◽  
JiXiang Liu ◽  
Stephen Yue ◽  
Jolene A. Bradford ◽  
William L. Godfrey
Keyword(s):  
Flow Cytometry ◽  
Bandpass Filter ◽  
Live Cell ◽  
Blue Dye ◽  
Dead Cell ◽  
Band Pass Filter ◽  
Scatter Plot ◽  
Pass Filter ◽  
Human Leukocytes ◽  
Polychromatic Flow Cytometry

Abstract Violet-excited fluorochromes are becoming more commonly used in polychromatic flow cytometry experiments. However, violet-excited fluorochromes with emissions longer than 450 nm have been shown to produce small signals relative to the autofluorescent background, usable only on densely expressed antigens, and are sometimes excited by a 488 nm argon ion laser. We have developed a novel violet-excited organic fluor, Pacific Orange™ dye, which has an emission maximum at 551 nm and which is not excited by 488 nm light. Pacific Orange dye is at least twice as bright as the other green emitting violet excitable dyes, Cascade Yellow™ dye and Alexa Fluor® 430 dye. Pacific Orange dye (585/42 nm bandpass filter) can be used for two color immunophenotyping with Pacific Blue ™ dye (450/50 nm band pass filter) with minimal compensation. Data is shown comparing a human CD4/CD8 combination using a direct antibody conjugate with a Zenon® labeling reagent bound to a primary antibody. CD45 antigen is easily resolved with Pacific Orange dye, allowing CD45/SSC gating of leukocytes using violet excitation. Pacific Orange and Pacific Blue dyes can be paired with the violet-excited Fixable Aqua dead cell stain (525/50 nm bandpass filter) to exclude dead cells from immunofluorescence staining. (Figure 1) Finally, a five-color human peripheral blood leukocyte panel is shown using only violet excitation, and pairing Pacific Orange anti-CD8 and Pacific Blue anti-CD4 with Qdot® 605, Qdot 655, and Qdot 705 nanocrystal streptavidin conjugates used sequentially with biotinylated anti-CD19, anti-CD3, and anti-CD56. (Figure 2) Pacific Orange dye provides a tool to transfer detection of abundant target antigens from 488 nm excitation to the violet laser, enabling the use the 488 laser for another marker. In addition, the use of multiple violet-excited dyes can enable the detectection of five or more additional markers to standard laser combinations for greater multiplexing in polychromatic flow cytometry. Figure 1. Immunophenotyping of mixed live and heat-killed human leukocytes using Pacific Orange dye, Pacific Blue dye and the Fixable Aqua dead cell reagent. Live cell events (Fixable Aqua stain-negative) were gated in the histogram (left) for display in the CD4/CD8 scatter plot (right). Figure 1. Immunophenotyping of mixed live and heat-killed human leukocytes using Pacific Orange dye, Pacific Blue dye and the Fixable Aqua dead cell reagent. Live cell events (Fixable Aqua stain-negative) were gated in the histogram (left) for display in the CD4/CD8 scatter plot (right). Figure 2. Five-color immunophenotyping of human leukocytes with Pacific Orange dye, Pacific Blue dye and three Qdot nanocrystal streptavidin conjugates using violet excitation. The Qdot nanocrystal staining was done with sequential staining and washing with biotinylated primary antibodies and streptavidin conjugates. Figure 2. Five-color immunophenotyping of human leukocytes with Pacific Orange dye, Pacific Blue dye and three Qdot nanocrystal streptavidin conjugates using violet excitation. The Qdot nanocrystal staining was done with sequential staining and washing with biotinylated primary antibodies and streptavidin conjugates.

scholarly journals Identification and Phenotyping of Leukocytes in Bovine Bronchoalveolar Lavage Fluid

2004 ◽  
Vol 11 (4) ◽  
pp. 795-798 ◽  
Author(s):  
E. C. Soethout ◽  
K. E. Müller ◽  
A. J. M. van den Belt ◽  
V. P. M. G. Rutten
Keyword(s):  
Flow Cytometry ◽  
Bronchoalveolar Lavage ◽  
Alveolar Macrophages ◽  
Bronchoalveolar Lavage Fluid ◽  
Lavage Fluid ◽  
Light Scatter ◽  
Specific Antibodies ◽  
Dual Laser

ABSTRACT A method is proposed to identify leukocyte subpopulations in bovine bronchoalveolar lavage fluid by dual-laser flow cytometry. The technique uses several parameters, i.e., exclusion of highly autofluorescent alveolar macrophages and inclusion of leukocytes on the basis of labeling by specific antibodies and light scatter characteristics.

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