Use of diploid and triploid trout erythrocytes as internal standards in flow cytometry

Pierrez, J.; Ronot, Xavier

doi:10.1002/cyto.990120311

Cited by 13 publications

(9 citation statements)

References 8 publications

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“…1 SD) for Scrippsiella hangoei flagellate cells and cysts from strain SHTV5 and the mixed-strain culture relative to triploid trout erythrocyte nuclei. Triploid trout unit 5 8.25 pg DNA per nucleus (Pierrez and Ronot 1991). ANKE KREMP AND MATTHEW W. PARROW Flagellate cells of S. hangoei occur in the Baltic Sea during winter and early spring, when the surface is typically ice covered and water temperatures are between 01 C and 41 C. When the surface ice melts and water temperatures begin to rise in late spring, encystment might be induced in natural populations of S. hangoei in a similar way as occurred in the culture experiments reported here.…”

Section: Discussionmentioning

confidence: 99%

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EVIDENCE FOR ASEXUAL RESTING CYSTS IN THE LIFE CYCLE OF THE MARINE PERIDINOID DINOFLAGELLATE,SCRIPPSIELLA HANGOEI¹

Kremp

Parrow

2006

Journal of Phycology

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Scrippsiella hangoei (Schiller) Larsen is a peridinoid dinoflagellate that grows during winter and spring in the Baltic Sea. In culture this species formed round, smooth cysts when strains were mixed, indicating heterothallic sexuality and hypnozygote production. However, cysts of the same morphology were also formed in clonal strains exposed to slightly elevated temperature. To better understand the role of cysts in the life cycle of S. hangoei, cyst formation and dormancy were examined in culture experiments and the cellular DNA content of flagellate cells and cysts was compared in clonal and mixed strains using flow cytometry. S. hangoei exhibited a high rate of cyst formation in culture. Cysts produced in both clonal and mixed strain cultures were thick-walled and underwent a dormancy period of 4 months before germinating. The S. hangoei flagellate cell population DNA distributions consisted of 1C, intermediate, and 2C DNA, indicative of respective eukaryotic cell cycle phases G1, S, and G2M. The majority (495%) of cysts had a measured DNA content equivalent to the lower 1C DNA value, indicating a haploid nuclear phase and an asexual mode of cyst formation. A small percentage (o5%) of cysts produced in the mixed strain culture had 2C DNA, and thus could have been diploid zygotes. These findings represent the first measurements of dinoflagellate resting cyst DNA content, and provide the first quantitative evidence for dinoflagellate asexual resting cysts. Asexual resting cysts may be a more common feature of dinoflagellate life cycles than previously thought.

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Section: Discussionmentioning

confidence: 99%

“… Measured 1C and 2C cellular DNA content estimates (means±1 SD) for Scrippsiella hangoei flagellate cells and cysts from strain SHTV5 and the mixed‐strain culture relative to triploid trout erythrocyte nuclei. Triploid trout unit=8.25 pg DNA per nucleus (Pierrez and Ronot 1991). …”

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confidence: 99%

EVIDENCE FOR ASEXUAL RESTING CYSTS IN THE LIFE CYCLE OF THE MARINE PERIDINOID DINOFLAGELLATE,SCRIPPSIELLA HANGOEI¹

Kremp

Parrow

2006

Journal of Phycology

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“…Previous research has demonstrated that polyploids possess larger erythrocytes (Licht and Bogart 1987;Boron 1994), and that the nuclear and cell size of erythrocytes uniformly increases with the level of ploidy (Small and Benfey 1987;Wolters et al 1982). Hence, the size of erythrocytes has been used as a criterion to identify ploidy (Small and Benfey 1987;Espinosa et al 2005;Harrell et al 1998;Thorgaard et al 1982;Pierrez and Ronot 1991;Cassani et al 1990). However, all these earlier data have been gained only from diploids and triploids, with no reports on higher ployploidy levels.…”

Section: Relationship Between Ploidy and Size Of Erythrocytesmentioning

confidence: 98%

Comparative study of erythrocytes of polyploid hybrids from various fish subfamily crossings

Liu

et al. 2009

Cell Tissue Res

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We have undertaken comparative studies of the number and phenotypes of erythrocytes in the peripheral blood of red crucian carp (RCC), blunt snout bream (BSB), and their hybrids, including triploids, tetraploids, and pentaploids. The results indicate that the mean nuclear volume of erythrocytes in peripheral blood increases regularly with increasing ploidy. Furthermore, many more mature erythrocytes have a dumbbell nucleus in the peripheral blood of polyploid hybrids compared with their diploid parents. With the increase in ploidy level, the percentage of such erythrocytes increases. The polyploid hybrids also have a large number of erythrocytes with abnormal shapes. For example, round and tear-shaped erythrocytes have been observed in the peripheral blood of polyploid hybrids. Since the erythrocytes in polyploid hybrids with their larger volume and lower specific surface area are unfavorable for the conveyance of oxygen, morphological variations of erythrocytes might improve defective blood circulation.

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“…These standards have a known DNA content and are typically used for calibrating flow cytometers. [28][29][30][31] They are used in this study to calibrate OPTM. In order to find an alternative stain to Feulgen, which provides stoichiometry as well as visual morphology in 3-D, we stoichiometrically optimized acidic thionin stain using different staining times.…”

Section: Introductionmentioning

confidence: 99%

Three-dimensional DNA image cytometry by optical projection tomographic microscopy for early cancer diagnosis

Agarwal

Biancardi

Patten³

et al. 2014

J. Med. Imag

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Abstract. Aneuploidy is typically assessed by flow cytometry (FCM) and image cytometry (ICM). We used optical projection tomographic microscopy (OPTM) for assessing cellular DNA content using absorption and fluorescence stains. OPTM combines some of the attributes of both FCM and ICM and generates isometric highresolution three-dimensional (3-D) images of single cells. Although the depth of field of the microscope objective was in the submicron range, it was extended by scanning the objective's focal plane. The extended depth of field image is similar to a projection in a conventional x-ray computed tomography. These projections were later reconstructed using computed tomography methods to form a 3-D image. We also present an automated method for 3-D nuclear segmentation. Nuclei of chicken, trout, and triploid trout erythrocyte were used to calibrate OPTM. Ratios of integrated optical densities extracted from 50 images of each standard were compared to ratios of DNA indices from FCM. A comparison of mean square errors with thionin, hematoxylin, Feulgen, and SYTOX green was done. Feulgen technique was preferred as it showed highest stoichiometry, least variance, and preserved nuclear morphology in 3-D. The addition of this quantitative biomarker could further strengthen existing classifiers and improve early diagnosis of cancer using 3-D microscopy.

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Use of diploid and triploid trout erythrocytes as internal standards in flow cytometry

Cited by 13 publications

References 8 publications

EVIDENCE FOR ASEXUAL RESTING CYSTS IN THE LIFE CYCLE OF THE MARINE PERIDINOID DINOFLAGELLATE,SCRIPPSIELLA HANGOEI¹

EVIDENCE FOR ASEXUAL RESTING CYSTS IN THE LIFE CYCLE OF THE MARINE PERIDINOID DINOFLAGELLATE,SCRIPPSIELLA HANGOEI¹

Comparative study of erythrocytes of polyploid hybrids from various fish subfamily crossings

Three-dimensional DNA image cytometry by optical projection tomographic microscopy for early cancer diagnosis

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Use of diploid and triploid trout erythrocytes as internal standards in flow cytometry

Cited by 13 publications

References 8 publications

EVIDENCE FOR ASEXUAL RESTING CYSTS IN THE LIFE CYCLE OF THE MARINE PERIDINOID DINOFLAGELLATE,SCRIPPSIELLA HANGOEI1

EVIDENCE FOR ASEXUAL RESTING CYSTS IN THE LIFE CYCLE OF THE MARINE PERIDINOID DINOFLAGELLATE,SCRIPPSIELLA HANGOEI1

Comparative study of erythrocytes of polyploid hybrids from various fish subfamily crossings

Three-dimensional DNA image cytometry by optical projection tomographic microscopy for early cancer diagnosis

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EVIDENCE FOR ASEXUAL RESTING CYSTS IN THE LIFE CYCLE OF THE MARINE PERIDINOID DINOFLAGELLATE,SCRIPPSIELLA HANGOEI¹

EVIDENCE FOR ASEXUAL RESTING CYSTS IN THE LIFE CYCLE OF THE MARINE PERIDINOID DINOFLAGELLATE,SCRIPPSIELLA HANGOEI¹