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PubMed1997

Flow cytometric analysis and confocal imaging of anticancer alkylaminoanthraquinones and their N-oxides in intact human cells using 647-nm krypton laser excitation.

Flow cytometry and laser-scanning confocal fluorescence microscopy have been used in the study of the pharmacodynamics, in single intact cells, of two novel alkylaminoanthraquinones (AQ4 and AQ6), structurally based upon the mid-red excitable but very weakly fluorescent anticancer agent mitoxantrone, together with their respective N-oxide derivatives (AQ4NO and AQ6NO). The drug design rationale was that N-oxide modifications generates prodrug forms suitable for selective bioreductive-activation in hypoxic tumor cells. DNA-binding ranked in the order of mitoxantrone > AQ6 > AQ4 > AQ6NO >> AQ4NO. Using both cytometric methods a similar ranking was found for whole cell and nuclear location in human transformed fibroblasts. However, AQ6 showed enhanced nuclear uptake compared with mitoxantrone, in keeping with its greater capacity to inhibit DNA synthesis. Partial charge neutralisation by N-oxide derivatization resulted in loss of DNA synthesis inhibition but retention of the ability to accumulate in the cytosol, an important property for prodrug development. We conclude that both flow cytometry and confocal imaging revealed biologically significant differences between analogues for subcellular distribution and retention properties. The study demonstrates the potential for these complementary 647-nm krypton laser line-based fluorometric methods to provide relevant structure-activity information in anthraquinone drug-design programmes.

Anthraquinones↗
PubMed1997

Sarcomatoid renal cell carcinoma: the chromophobe connection.

Eleven cases of sarcomatoid renal cell carcinoma were studied to determine the relative frequency of various subtypes of renal cell carcinoma that may be associated with sarcomatoid transformation. The epithelial components in these tumors were subcategorized according to established histologic criteria into chromophobe carcinoma (n = 6 cases), clear cell carcinoma (n = 3), papillary carcinoma (n = 1), and indeterminate (n = 1). In nine cases, material was available for immunohistochemical and DNA ploidy studies. The sarcomatoid component in all cases showed positivity for epithelial membrane antigen cytokeratin, indicating an epithelial derivation of these cells. Staining for mesenchymal markers was mostly negative, except for vimentin, which reacted strongly in all cases. DNA ploidy studies using flow cytometry and cell image analysis provided very similar results. Five of five chromophobe sarcomatoid carcinomas showed hypodiploid cell lines in the epithelial areas, whereas the sarcomatoid components mostly showed aneuploid peaks. In the remaining cases, DNA ploidy pattern was more variable. These findings indicate that chromophobe carcinoma may be the most frequent epithelial component associated with sarcomatoid renal cell carcinoma.

Adult↗
PubMed1989

The role of cell cycle activity in the generation of morphologic heterogeneity in non-Hodgkin's lymphoma.

Cellular and nuclear size heterogeneity are frequently observed in non-Hodgkin's lymphomas, yet the biological basis of this morphologic variability is not well understood. The possible relationship between cell size and cell cycle activity in malignant lymphomas was investigated using multiparameter flow cytometry and digital image analysis. Flow cytometric analysis of ten cases of B cell lymphoma revealed that, regardless of the diagnosis or the proportion of large cells present, large neoplastic cells showed a much greater proportion of S + G2/M phase activity (mean, 35.2%; range, 14.6% to 70.5%) than did small cells (mean, 4.4%; range, 0.5% to 18.4%). Further studies aimed at distinguishing the features of cycling and noncycling cells were performed using quantitative image analysis of nuclear staining by Ki-67 antibody. In both benign and malignant lymphoid infiltrates, resting (Ki-67-negative) nuclei were uniformly smaller than Ki-67-positive nuclei and exhibited a restricted size distribution, whereas the Ki-67-positive nuclei had a broad size distribution and included the vast majority of large forms. Thus, the actively cycling component of the tumor consisted of cells with large nuclei. These studies support the hypothesis that cellular and nuclear size heterogeneity in malignant lymphomas is related to cell cycle phase. The combined influences of cell differentiation and proliferation on cellular morphology can explain many of the histologic features observed in non-Hodgkin's lymphomas.

Antibodies, Monoclonal↗
PubMedDate not supplied

[Methods of automated cell analysis and their application in radiation biology].

The present review is concerned with the methods of automated analysis of biological micro-objects and covers two groups into which all the systems of automated analysis can be divided--systems of flow (flow cytometry) and scanning (image analysis systems) type. Particular emphasis has been placed on their use in radiobiological studies, namely, in the micronucleus test, a cytogenetic assay for monitoring the clastogenic action of ionizing radiation commonly used at present. It is evident that the problem is acute, with of radiobiologists' interest in the biological action of low-dose radiation recently increasing. In addition, the estimation of a low-level damage requires the analysis of a large number of experimental objects. Examples of using both the methods elsewhere and actual setups are given. The analysis of advantages and disadvantages of the methods of automated cell analysis enables us to choose more thoroughly between the systems of flow and scanning type to use them in particular research.

Animals↗
PubMed1993

Ploidy changes between diagnosis and relapse in childhood renal tumours.

Ploidy patterns, analysed by flow cytometry (FCM) and image analysis (IA), were investigated at relapse in a group of six children with renal tumours [five Wilms' tumour (WT) and one bone metastasizing renal tumour of childhood (BMRTC)] and results compared with diagnostic profiles. IA detected one or more aneuploid populations in five of 12 tumours which were diploid on FCM. Patterns in three of six patients [two with unfavourable histology (UH) and one with favourable histology (FH)] were aneuploid at diagnosis and relapse, two patients (one FH, one BMRTC) developed aneuploid features at relapse and one patient with a tetraploid tumour was diploid at relapse. Histology patterns were similar at diagnosis and relapse in all patients. Three of six patients (two UH, one BMRTC) have died of disease. This report highlights (1) the superiority of IA over FCM in detecting aneuploid populations and (2) changes in ploidy status which have not previously been reported in these tumours. Overinterpretation of DNA status at relapse may prove misleading.

Aneuploidy↗
PubMed1996

Stimulation of calcium uptake in cultured rat hippocampal neurons by 2,3,7,8-tetrachlorodibenzo-p-dioxin.

This study examined the effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD) and related compounds on the uptake of intracellular calcium ([Ca2+]i) in primary cultures of rat hippocampal neuronal cells. [Ca2+]i levels were detected and quantified by interactive laser cytometry with microscopic image analysis. Cells were noninvasively labeled with fluo-3/AM and all experiments were conducted on cultured rat hippocampal neurons 14 days in culture. Treatment of cell cultures with 2,3,7,8-TCDD (10-100 nM) resulted in a rapid concentration-dependent increase in [Ca2+]i associated with a decrease in mitochondrial membrane potential and activation of alpha-protein kinase C (alpha-PKC). In contrast, 1,2,3,4-TCDD, a weak Ah receptor agonist, had no effect on [Ca2+]i at concentrations as high as 10 microM and similar results were also observed for 2,2',5,5'-tetrachlorobiphenyl. Maximal [Ca2+]i was observed within 30 s after addition of 2,3,7,8-TCDD and remained elevated (at higher concentrations) above resting levels for the duration of the experiment. This rapid increase in [Ca2+]i was blocked by addition of EDTA (2 mM) to the external medium or by pretreatment of the cells with the calcium channel antagonist nifedipine (10 microM). However, pretreatment of the cells with 100 microM cycloheximide failed to block calcium uptake in neuronal cells. These data indicate that rat hippocampal neuronal cells are responsive to 2,3,7,8-TCDD; however, the mechanism is not associated with altered gene transcription and may involve cellular targets.

Analysis of Variance↗
PubMed1997

Image cytometric nuclear DNA quantitation of paragangliomas in tissue sections. Prognostic significance.

OBJECTIVE: Image cytometric quantitation of nuclear DNA of paragangliomas may provide prognostic information that cannot be obtained from histopathologic study. Flow cytometry has demonstrated DNA aneuploid tumors to have a higher risk of progression than diploid neoplasms. STUDY DESIGN: DNA ploidy of 56 paragangliomas was assessed by image cytometry of 5-micron, Feulgen-stained, formalin-fixed, paraffin-embedded tissue sections. RESULTS: Thirty-three (59%) paragangliomas were diploid and 23 (41%) aneuploid. Of the 30 adrenal pheochromocytomas, 15 (50%) were diploid. Thirteen (93%) of the 14 carotid body tumors were diploid. Five of seven (71%) glomus jugulare tumors and two of five (40%) extraadrenal paragangliomas were aneuploid. During a mean follow-up of 57 months (range, 1 month to 36 years) of 44 patients with 47 paragangliomas, 33 (75%) were alive and without disease; 7 (16%), including 1 glomus jugulare, 2 carotid body and 4 pheochromocytoma patients, developed recurrences/metastases. By multivariate analysis, image cytometric DNA ploidy was predictive of disease-free survival for adrenal pheochromocytomas. No significant differences in overall survival, disease-free survival or recurrence/metastasis rate were noted between other diploid and aneuploid tumors. CONCLUSION: Aneuploidy suggests a risk of early recurrence for adrenal pheochromocytomas.

Adrenal Gland Neoplasms↗
PubMed1995

Role of the pathologist in biomarker studies.

Cancer chemoprevention is defined as intervention by chemical agents prior to invasion to inhibit or slow the carcinogenic process. Using surrogate endpoint biomarkers in chemoprevention studies may reduce the size, length and cost of clinical prospective randomized trials in high-risk populations. Intermediate biomarkers are measurable alterations in the tissues at risk and include differentiation, genetic composition, biochemical expression, and proliferation. Assessment is possible because invasive epithelial neoplasms are known to begin as intraepithelial proliferations with a spectrum of cellular abnormalities extending to carcinoma in situ. Genetic heterogeneity begins in the intraepithelial phase; a stochastic accumulation of genetic errors characterizes the progression of clonal evolution within the tumor through the process of invasion and metastasis. Pathologic features associated with this process include tumor classification as well as whether it is intraepithelial or invasive. If the process is intraepithelial, the grade and extent of the intraepithelial lesion are reported. If the neoplasm is invasive, tumor size, extent, degree of differentiation (histologic and nuclear grade), mitotic rate, vascular invasion, and lymph node involvement are evaluated. In assessing biomarkers relevant chemoprevention, and without complete regression of the neoplasm with the chemopreventive agent or agents, measurable parameters along with histopathologic features are applicable. Three methods readily applicable for this purpose that can be applied to paraffin-embedded, formalin-fixed tissue include quantitative pathology, immunohistochemistry, and molecular biologic applications. These methods require some consistency in handling and processing the tissues under study; results may deteriorate due to a number of processing variables, including time to fixation, time in fixative, and fixative type. Quantitative pathology, including static image analysis and flow cytometry, can determine total DNA content. Using static image analysis, very small tumors can be studied. In addition, adjacent intraepithelial and invasive components of a tumor may be studied from a single slide. Steroid receptors, oncogenes, and other proteins detectable through immunohistochemical or molecular biologic methods can be quantitated by this technique as well. Cell cycle synthetic function is assayable by both methods. Flow cytometry can calculate the total percentage of cells in S-phase, or the tumor cell S-phase fraction based on the percentage of cells detected between the G0, G1 peak and the G2 + M peak. A similar approach is generally not applicable with current image analysis equipment; however, cell cycle related proteins such as MIB-1 (Ki-67 associated) can be quantified. Immunohistochemical methods can employ a wide variety of monoclonal antibodies to detect oncogene related proteins, including HER-2/neu (c-erbB-2) and p53. Molecular biologic methods, including in situ hybridization, polymerase chain reaction, and in situ PCR, can have many applications when applied to paraffin-embedded tissues, including detection of viral DNA, identification and measurement of apoptosis, and defining gene deletions.

Biomarkers, Tumor↗
PubMed2006

Novel aspects of systems biology and clinical cytomics.

The area of Cytomics and Systems Biology became of great impact during the last years. In some fields of the leading cytometric techniques it represents the cutting edge today. Many different applications/variations of multicolor staining were developed for flow- or slide-based cytometric analysis of suspensions and sections to whole animal analysis. Multispectral optical imaging can be used for studying immunological and tumorigenic processes. New methods resulted in the establishment of lipidomics as the systemic research of lipids and their behavior. All of these development push the systemic approach of the analysis of biological specimens to enhance the outcome in the clinic and in drug discovery programs.

Biomedical Research↗
PubMed1991

Correlation of DNA ploidy with c-erbB-2 expression in preinvasive and invasive breast tumors.

Detection of c-erbB-2 oncogene product expression by monoclonal antibody staining (avidin-biotin technique) in formalin-fixed, paraffin-embedded atypical hyperplasias (AH, n = 20), intraductal carcinomas (IDCA, n = 27) and invasive carcinomas (INVCA, n = 48) was compared to ploidy determinations obtained by flow cytometry (INVCA) or image analysis (AH, IDCA). Cytoplasmic membrane staining was present in 11/48 (23%) INVCA and 8/27 (30%) IDCA but none of the AH. Tumors with abnormal DNA content expressed c-erbB-2 more frequently: INVCA, 2/19 (11%) diploid range versus 9/29 (31%) aneuploid; IDCA, 1/7 (14%) diploid range versus 7/20 (35%) aneuploid. Poorly differentiated (nuclear grade) IDCA or INVCA were also more frequently stained (14/35, 40%) than were well or moderately differentiated cases (5/40, 12.5%). Oncogene product expression and DNA content derangements may be related biologic parameters in breast neoplasia, and both are highly associated with cytologic nuclear abnormalities.

Adenocarcinoma↗
PubMed1994

Surface expression and distribution of Fc receptor III (CD16 molecule) on human natural killer cells and polymorphonuclear neutrophils.

Human natural killer cells and polymorphonuclear neutrophils constitutively express the low-affinity IgG Fc receptor (Fc gamma RIII, CD16 molecule). To investigate cell surface morphology, antigenic receptor density, and topographical distribution of Fc gamma RIII on the plasma membrane of natural killer cells and polymorphonuclear neutrophils, conventional scanning electron microscopy (SEM), flow cytometry, and immunoscanning electron microscopy were used. Fc gamma RIII was detected with an indirect immunogold labeling procedure, and receptors were visualized in the backscattered and secondary electron imaging mode of SEM. Natural killer cells showed a cell surface morphology compatible with lymphocytic differentiation characterized by microvilli and microridges. Polymorphonuclear neutrophils showed surface features characterized by ridges with folds and scattered short microvilli. Natural killer cells displayed a lower cell labeling density, whereas polymorphonuclear neutrophils showed a high level of expression of Fc gamma RIII on the plasma membrane by quantitative analysis with SEM in the backscattered electron imaging mode. Flow cytometry analysis confirmed these findings. Analysis of the topographical distribution of Fc gamma RIII antigenic receptor sites by SEM in the backscattered and secondary electron imaging modes showed that Fc gamma RIII on natural killer cells are randomly distributed, whereas Fc gamma RIII are located on ridges and folds of the plasma membrane of polymorphonuclear neutrophils. These observations suggest that natural killer cells and polymorphonuclear neutrophils differ in their levels of expression and topographic distribution of Fc gamma RIII on the plasma membrane. This different spatial distribution of Fc gamma RIII would provide morphological evidence of certain cellular functions mediated by natural killer cells and polymorphonuclear neutrophils.

Cell Count↗
PubMed2004

Mutation in the leucine-rich repeat C-flanking region of platelet glycoprotein Ib beta impairs assembly of von Willebrand factor receptor.

We describe a syndrome of thrombocytopenia, bleeding episodes, congenital heart disease and facial dysmorphism in a newborn infant, and trace the cause to mutations on chromosome 22 that involve the gene for platelet glycoprotein Ib beta (GPIb beta, Human Genome Organisation gene symbol GPIBB), a critical component of the von Willebrand factor (vWF) receptor. Fluorescence in situ hybridization in transformed lymphoblasts revealed hemizygous microdeletion of 22q11.2 containing the GP1BB locus. DNA sequencing revealed a C to T transition in the patient's remaining GP1BB allele, predicting a novel proline to serine substitution (Pro96Ser) in the carboxyterminal flanking domain of a leucine-rich repeat. We characterized the mutant GP1BB allele by expression in a cell line (CHO alpha IX) that stably expresses two other components of the vWF receptor, GPIb alpha and GPIX. Flow cytometry and confocal imaging of transfected CHO alpha IX cells demonstrated that P96S GPIb beta abrogates surface assembly of the complex, consistent with platelet flow cytometry studies in the patient. Based on sequence homology to the known crystal structures of two other leucine-rich repeat proteins, the human Nogo receptor and GPIb alpha, we propose a new structural model of GPIb beta. The model refutes earlier assumptions about cysteine-cysteine interactions in the amino-terminal region of GPIb beta, and predicts a hydrophobic patch the burial of which may contribute to proper conformation of the fully assembled vWF receptor complex.

Abnormalities, Multiple↗
PubMed2002

HLA class I expression in primary hepatocellular carcinoma.

AIM: To investigate whether CTL vaccine therapy is suitable for primary hepatocellular carcinoma (HCC) from the viewpoint of HLA class I antigens expression. METHODS: The immunocytochemistry, image analysis, flow cytometry, and labeled streptavidin biotin (LSAB) method of immunohistochemistry were applied respectively to study 4 HCC cell lines (e.g. Alexander, HepG2, SMMC-7721, and QGY-7703) cultured in vitro and 6 frozen tissue specimens of HCC. RESULTS: The positive control cell line Raji had very strong positive staining. Most mitotic and nonmitotic cells of the 4 HCC cell lines had various intensity of HLA class I antigens expression. The negative control cell K562 and the control slides of all the cell lines had no positive staining. In the 6 HCC specimens immunohistochemically studied, histological normal hepatocytes had no or very weak positive staining and the liver sinus had very strong positive staining. Most HCC cells in the sections from the 6 HCC specimens had strong positive HLA classIantigens staining. The positive staining was located in the cytoplasm, the perinuclear area, and at the cell membrane of the liver cancer cells. Flow cytometry also revealed that Raji and those 4 HCC cell lines had strong HLA classIantigens expression, which was confirmed quantitatively by the image analysis. It showed that the objective grayscale values of Raji and those 4 HCC cell lines were significantly different from that of K562 (Raji 114.04+/-10.94, Alexander 165.97+/-5.35, HepG2 167.02+/-12.60, QGY-7703 161.46+/-7.13, SMMC-7721 165.93+/-5.21, K562 244.89+/-4.60, P<0.01). Significant differences were also found between Raji and the 4 HCC cell lines. CONCLUSION: HCC cells express HLA class I antigens strongly. From this point of view, the active specific immunotherapy of CTL vaccine is suitable and practicable for HCC.

Cancer Vaccines↗
PubMed1997

Prognostic value of S-phase fraction in lymph-node-negative breast cancer by image and flow cytometric analysis.

Cellular DNA content and proliferation rates have been suggested as prognostic factors in breast carcinomas. A series of 271 lymph-node negative breast carcinoma patients without adjuvant therapy was reviewed (mean follow-up, 108 mo). Tumor cells from the same paraffin-embedded block tissue (Hedley's method) were analyzed by image analysis (IA) in Feulgen-stained smears and by flow cytometric analysis (FC). Clinicopathologic features, ploidy, S-phase fraction, and percentage of tumor cells with more than 5n DNA content (in diploid tumors, by IA) were related to outcome. The results of IA and FC were compared in 115 cases. Tumor size, histologic grade, desmoplasia and S-phase fraction were significant predictors of survival in multivariate analysis (Cox proportionate regression) (P < or = 0.03). Ploidy by the two methods showed agreement in 100 carcinomas (87%). Of the 15 discordant cases, FC detected 6 multiploid and 4 aneuploid-peridiploid. In contrast, IA detected more tetraploid carcinomas. Tumor size, histologic grade, desmoplasia, and S-phase fraction were independent predictors of long-term prognosis in our patients. Ploidy and percentage of tumor cells with more than 5n DNA content were not prognostic indicators. FC detected aneuploidy more frequently than did IA.

Adult↗
PubMedDate not supplied

Proliferative activity in invasive breast carcinoma: a comprehensive comparison of MIB-1 immunocytochemical staining in aspiration biopsies to image analytic, flow cytometric and histologic parameters.

OBJECTIVE: To use MIB-1 antibody to assess proliferative activity in fine needle aspiration (FNA) samples of invasive breast carcinoma and compare these results to multiple other measures of proliferative activity. STUDY DESIGN: FNA slides from 62 patients with invasive breast carcinoma were subjected to staining with MIB-1. Quantitative MIB-1 values were compared to image analytic proliferative fractions (IPF) obtained from the same FNAs. MIB-1 values were also compared to flow cytometric S-phase fractions (SPF) and S + G2/M-phase fractions (FPF) and to histologic assessment of mitotic count (MC) in resected tumors. RESULTS: MIB-1 values, IPF, SPF, FPF and MC were suitable for evaluation in 55, 53, 50, 50 and 56 cases, respectively. MIB-1 values showed good correlation with IPF in FNAs (correlation coefficient = .57, P <.00001). MIB-1 values also showed correlation with SPF (correlation coefficient =.447, P = .003), FPF (correlation coefficient = .325, P = .023) and MC (correlation coefficient = .402, P = .01) in resected tumors. CONCLUSION: This study supports the use of MIB-1 values obtained from FNA samples for assessment of proliferative activity in invasive breast carcinoma, based on correlation of these values with multiple other parameters of proliferative activity. Assessment of these values can play a role in predicting prognosis and in selecting patients with invasive carcinoma of the breast for preoperative or adjuvant chemotherapy.

Adenocarcinoma↗
PubMed1999

[Bladder transitional cell carcinoma rating, systematization of grading and value of DNA ploidy].

UNLABELLED: TCCs are an heterogenous group of tumors with an uncertain biologic behaviour, especially intermediate grade (G2). Histologic grade and pathologic stage have revealed only partially useful on predicting the outcome. OBJECTIVE: Search and statement of objective and cuantitative parameters able to define prognostic subgroups to TCCs. MATERIAL AND METHOD: We studied consecutive 106 patients with TCC in order to: 1) Identify every variable with independent predictive value in classifying cases of TCC in a three (1,2,3) or two (high/low) grades systems. 2) Make a correlation between DNA ploidy obtained by image analysis and DNA ploidy and S-phase obtained by flow cytometry. RESULTS: Every variable used to define the histologic grade showed significative correlation with both gradation systems. In multivariate analysis, the presence of superficial cells and the mitotic counts revealed us the most valuable variables in predicting the histologic grade. DNA-ploidy (both obtained by static an flow cytometry) correlated well with grade, stage, growth pattern and necrosis, whereas S-phase did so with grade, mitotic index and DNA ploidy. Image cytometry showed similars results to those of flow cytometry and also was able to detect aneuploidy when an situ carcinoma or dysplasia were present in the adyacent mucosa.

Carcinoma, Transitional Cell↗
PubMed2002

The use of field emission scanning electron microscopy to assess recombinant adenovirus stability.

A field emission scanning electron microscopy (FESEM) method was developed to assess the stability of a recombinant adenovirus (rAd). This method was designed to simultaneously sort, count, and size the total number of rAd viral species observed within an image field. To test the method, a preparation of p53 transgene-expressing recombinant adenovirus (rAd/p53) was incubated at 37 degrees C and the viral particles were evaluated by number, structure, and degree of aggregation as a function of time. Transmission electron microscopy (TEM) was also used to obtain ultrastructural detail. In addition, the infectious activity of the incubated rAd/p53 samples was determined using flow cytometry. FESEM image-analysis revealed that incubation at 37 degrees C resulted in a time-dependent decrease in the total number of detectable single rAd/p53 virus particles and an increase in apparent aggregates composed of more than three adenovirus particles. There was also an observed decrease in both the diameter and perimeter of the single rAd/p53 viral particles. TEM further revealed the accumulation of damaged single particles with time at 37 degrees C. The results of this study demonstrate that FESEM, coupled with sophisticated image analysis, may be an important tool in quantifying the distribution of aggregated species and assessing the overall stability of rAd samples.

Adenoviruses, Human↗
PubMed1994

Cellular effects of image diagnostic ultrasound on murine spermatogenesis monitored by flow cytometry.

A study has been carried out to evaluate the possible cellular effects induced by image diagnostic ultrasound on murine spermatogenetic cells. Exposure to ultrasound was carried out using a commercial diagnostic instrument that operates in B-mode. Male hybrid F1 mice, aged 8-10 weeks, were exposed to ultrasound for 30 min and observed from 7 to 35 days after treatment. Flow cytometric analysis has been used to monitor the relative frequency of the different types of spermatogenetic cells. This analytical approach showed changes in cell frequency in the compartment containing elongated spermatids which was used as an endpoint. A statistically significant decrease in the frequency of this cell type was observed 21, 28 and 35 days after exposure. These changes suggest that there may be a cytotoxic and/or cytostatic effect on spermatocytes and spermatogonia. These results showed that image diagnostic ultrasound induces effects on murine spermatogenesis at cellular level and that the flow cytometric approach makes it possible to identify quantitative cellular changes with reference to specific cell type.

Animals↗

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