scholarly journals Formalin Versus Bouin Solution for Testis Biopsies: Which Is the Better Fixative?

2020 ◽  
Vol 13 ◽  
pp. 2632010X1989726 ◽  
Author(s):  
James L Ellenburg ◽  
Peter Kolettis ◽  
Joseph C Drwiega ◽  
Anna M Posey ◽  
Matthew Goldberg ◽  
...  
Keyword(s):  
Basement Membrane ◽  
Nuclear Membrane ◽  
Cytoplasmic Membrane ◽  
Testicular Tissue ◽  
Seminiferous Tubules ◽  
High Quality ◽  
Fixed Tissue ◽  
Formalin Fixed Tissue ◽  
Histologic Evaluation

Purpose: Some resources recommended Bouin solution for the fixation of testis biopsy specimens. We compared the histologic quality of rat testicular tissue using buffered formalin and Bouin solution as fixatives. Methods: We prospectively compared the histologic quality of rat testicular tissue fixed in Bouin solution versus formalin. Testicular tissue was harvested post-mortem from six rats. Each testis was removed and sectioned in half; one half was fixed in formalin and one half in Bouin solution. Testicular tissue histology (nuclear membrane detail, nuclear granularity, cytoplasmic granularity, cytoplasmic membrane detail, and basement membrane detail) was graded as high quality (2) or low quality (1). Sloughing of cells into the lumens of the seminiferous tubules was graded on a 0-3 scale (0=none, 1=mild, 2=moderate, 3=extensive). Results: All slides regardless of fixative were of appropriate quality for the histologic evaluation of spermatogenesis. The average sloughing score for formalin cases was 1.4 and for Bouin cases 1.6. Formalin fixed tissue was found to have high quality nuclear membrane detail (2), nuclear granularity (1.9), and basement membrane detail (2). Cytoplasmic granularity was of lesser but adequate quality (1.4). Cytoplasmic membrane detail was poor, (1). Tissue fixed with Bouin solution had high quality basement membrane detail (2) and adequate cytoplasmic granularity (1.5), nuclear membrane detail (1.3) and nuclear granularity (1.4). Cytoplasmic membrane detail was poor (1). Conclusion: Compared to Bouin solution, formalin fixation of rat testicular tissue produced adequate histology for the evaluation of spermatogenesis and may be superior to Bouin solution for certain cytologic features.

scholarly journals Effects of microgravity or simulated launch on testicular function in rats

1992 ◽  
Vol 73 (2) ◽  
pp. S174-S185 ◽  
Author(s):  
R. P. Amann ◽  
D. R. Deaver ◽  
B. R. Zirkin ◽  
G. S. Grills ◽  
W. J. Sapp ◽  
...  
Keyword(s):  
Germ Cells ◽  
Quantitative Methods ◽  
Smooth Endoplasmic Reticulum ◽  
Testicular Tissue ◽  
Seminiferous Tubules ◽  
Specific Gene ◽  
Fixed Tissue ◽  
Treatment Groups ◽  
Essentially Normal ◽  
Peripheral Blood Plasma

Testes from flight rats on COSMOS 2044 and simulated-launch, vivarium, or caudal-elevation control rats (5/group) were analyzed by subjective and quantitative methods. On the basis of observations of fixed tissue, it was evident that some rats had testicular abnormalities unassociated with treatment and probably existing when they were assigned randomly to the four treatment groups. Considering rats without preexisting abnormalities, diameter of seminiferous tubules and numbers of germ cells per tubule cross section were lower (P less than 0.05) in flight than in simulated-launch or vivarium rats. However, ratios of germ cells to each other or to Sertoli cells and number of homogenization-resistant spermatids did not differ from values for simulated-launch or vivarium controls. Expression of testis-specific gene products was not greatly altered by flight. Furthermore, there was no evidence for production of stress-inducible transcripts of the hsp70 or hsp90 genes. Concentration of receptors for rat luteinizing hormone in testicular tissue and surface density of smooth endoplasmic reticulum in Leydig cells were similar in flight and simulated-launch rats. However, concentrations of testosterone in testicular tissue or peripheral blood plasma were reduced (P less than 0.05) in flight rats to less than 20% of values for simulated-launch or vivarium controls. Thus spermatogenesis was essentially normal in flight rats, but production of testosterone was severely depressed. Exposure to microgravity for greater than 2 wk might result in additional changes. Sequelae of reduced androgen production associated with microgravity on turnover of muscle and bone should be considered.

Improvement of the Quality of Frozen Sections from Formalin fixed Tissue

1990 ◽  
Vol 65 (1) ◽  
pp. 43-44
Author(s):  
Toshihiro Ishii ◽  
Kumiko Kasama ◽  
Mayumi Kondo
Keyword(s):  
Frozen Sections ◽  
Fixed Tissue ◽  
Formalin Fixed Tissue ◽  
Formalin Fixed

25 Cryopreservation of horse testicular tissue as a model for rhinoceros

2019 ◽  
Vol 31 (1) ◽  
pp. 138 ◽  
Author(s):  
M. C. Gómez ◽  
A. Alrashed ◽  
C.-Y. Su ◽  
B. Durrant
Keyword(s):  
Basement Membrane ◽  
Dimethyl Sulfoxide ◽  
Structural Integrity ◽  
Membrane Damage ◽  
Genetic Material ◽  
Testicular Tissue ◽  
Live Cells ◽  
Seminiferous Tubules ◽  
Positive Interaction ◽  
Cryoprotectant Solution

Cryopreservation of testicular tissue (TT) allows retention of valuable genetic material that can be used for conservation of endangered species, such as the northern white rhinoceros (NWR; Ceratotherium simum cottoni). Previously, we found that cryopreservation of NWR TT with a slow controlled cooling rate (CR) method induced morphological alterations in the seminiferous tubules (ST). However, the relative influence of CR, type of medium, and condition of TT from the aged NWR male on TT integrity was not clear. Due to the limited availability of rhinoceros TT, we used the horse as a model for optimization of TT cryopreservation. We evaluated the effect of (1) cryoprotectant solution [PBS (PBS +1.5M dimethyl sulfoxide) v. DMEM (DMEM/F12+10.0% fetal bovine serum+0.05M sucrose+1.5M dimethyl sulfoxide)] and (2) CR [CR1 (−2.0°C min−1 from 0°C to −4.0°C, −15°C min−1 to −12°C, and −0.3°C min−1 to −40°C in a programmable freezer) v. CR2 (same as CR1 but cooled to −8°C and held for 5min before cooling to −40°C) v. CR3 (−1.0°C min−1 from 0°C to −80°C in a CoolCell® freezing device; Corning, Corning, NY, USA)] on the structural integrity of ST from a 2-year-old horse (n=20 ST), cell viability, and expression of spermatogonial stem cells (SSC; GFRα1, and GRP125) and pluripotent markers (SSEA-4, SSEA-1, and OCT-4) in spermatogonial cells isolated from TT frozen with the above treatments (n=3). We found a positive interaction between CR and cryoprotectant solution on structural integrity of fixed and stained TT after freezing in PBS and CR2 that resulted in lower detachment of epithelium cells from the basement membrane (score±standard error of the mean; 0.50±0.1) than that of TT frozen in PBS and CR1 and CR3 (1.00±0.1 and 1.80±0.1, respectively; P<0.001) or in DMEM and CR1 (1.25±0.1), CR2 (1.35±0.1), and CR3 (1.40±0.1; P<0.01) and in lower incidence of basement membrane damage (0.75±0.1) than that of TT frozen in PBS and CR1 (1.17±0.07) and CR3 (1.16±0.07) or in DMEM and CR1 (1.10±0.1), CR2 (1.15±0.1), or CR3 (1.45±0.1; P<0.01). A lower rate of pyknosis was observed in TT frozen with PBS (1.15±0.06) than in TT frozen in DMEM (1.43±0.06; P<0.001). Overall, integrity of ST was improved when TT was frozen in PBS at CR2 having similar percentages of ST with intact epithelium (60%) and basement membrane (35%) as that of refrigerated TT (45 and 50%, respectively) but different from that of TT frozen with PBS at CR1 (10 and 15%, respectively; P<0.05). Flow cytometry analysis of spermatogonial cells revealed that the percentages of live cells from TT frozen in PBS (CR1: 61.5±7.4%; CR2: 59.7±4.8%; CR3: 51.5±4.1%) or DMEM (CR1: 66.2±6.0%; CR2: 59.8±6.0%; CR3: 58.9±6.9%), and expression of SSC and pluripotent markers was similar among all freezing treatments. However, the percentages of live cells from frozen-thawed TT were lower than those of cells isolated from refrigerated TT (80.6±2.2%; P<0.001). Overall, our results showed that (1) structural integrity of horse ST was better maintained when TT was frozen in PBS at CR2 and (2) SSC can be isolated from frozen-thawed TT with a similar relative frequency to that of refrigerated TT.

187 Morphological Appearance and Expression of Spermatogonial Stem Cell Markers in White Rhinoceros Testicular Tissue

2018 ◽  
Vol 30 (1) ◽  
pp. 233
Author(s):  
M. C. Gomez ◽  
Y. Cates ◽  
D. B. Stansfield ◽  
C. Young ◽  
R. Klee ◽  
...  
Keyword(s):  
Basement Membrane ◽  
Mammalian Species ◽  
Testicular Tissue ◽  
Seminiferous Tubules ◽  
Stem Cell Markers ◽  
Tissue Preparation ◽  
Fresh Tissue ◽  
Morphological Alterations ◽  
White Rhinoceros

Spermatogonial stem cells (SSC) have been isolated from testicular tissue (TT) of several mammalian species and differentiated into mature spermatozoa following transplantation or in vitro culture. The northern white rhinoceros (NWR; Ceratotherium simum cottoni) is critically endangered. Thus, frozen NWR TT, cryopreserved and stored at the San Diego Zoo’s Frozen Zoo®, potentially contain SSC that could be a source of spermatozoa. The method used for cryopreserving TT may affect the integrity and number of SSC. Therefore, identifying alterations in the seminiferous tubules (ST) of frozen-ndash;thawed-NWR TT will provide insight into the condition of the SSC. Therefore, our aims were to (1) determine the effect of freezing rhinoceros TT on the structure of epithelium, and (2) identify SSC (GFRα1, GPR125) and pluripotent (SSEA-4 and Oct-4) markers. Testicular tissue of an adult NWR and a stillborn southern white rhinoceros (SWR) were frozen by equilibration of TT for 30 min at 4.0°C in PBS and 1.5 M dimethyl sulfoxide (DMSO), cooled at 2.0°C/min to −4.0°C, 0.3°C/min to −40°C, and plunged into liquid nitrogen. Tissues were thawed at 37°C in a water bath and DMSO removed in a 4-step dilution. Tissue was then fixed, dehydrated, and paraffin embedded. For morphological evaluations, frozen-ndash;thawed tissue was sectioned and stained with hematoxylin and eosin (H&E). The TT from both rhinoceros collected immediately after death (fresh) and stained with H&E were used as a control for cryopreservation. Localization of SSC and pluripotent markers in ST of frozen-ndash;thawed TT was detected by immunohistochemistry. Morphologically, fresh-NWR TT was severely altered, displaying large epithelium gaps and partial (62.2%) or total detachment (37.7%) from, and slight damage (35.5%) to, the basement membrane. The number of pyknotic nuclei per ST was moderate (15.6 ± 7.2%). Many of these changes could have resulted from autolysis and handling before tissue preparation. In contrast, histological appearance of fresh-SWR was good, with 98.3% of the tubules intact, and a small proportion of pyknotic cells (0.8 ± 1.5%). Seminferous tubule (n = 30/male) length and width (μm; ± SEM) differed between NWR (635.2 ± 34.4 × 214.6 ± 10.8) and SWR (277.7 ± 13.8 × 73.2 ± 2.4; P < 0.05). Damages after cryopreservation compared with fresh tissue comprised (1) epithelium detachment, NWR = 100% (P < 0.0001), and SWR = 43.3% (P < 0.001); (2) basement membrane alteration, only in NWR (93.0%; P < 0.001); and (3) decreased length and width in the ST, NWR = 409.4 ± 18.1 × 173.4 ± 8.2 (P < 0.05), and SWR = 195.2 ± 8.3 × 61.6 ± 2.8 (P < 0.05), with loss of lumen in both males. Immunohistochemistry revealed that NWR expressed GFRα1 and GPR125 at various stages of spermatogenqaesis, whereas Oct-4 was detected in few cells. In contrast to NWR, Oct-4 expression in SWR was located at the basement membrane; SSEA-4 was not detected in either male. In conclusion, freezing-induced morphological alterations in rhinoceros ST and positive expression of markers for SSC and pluripotency suggest the presence of SSC. Further studies are required to evaluate the viability of rhinoceros SSC.

scholarly journals Thin basement membrane nephropathy cannot be diagnosed reliably in deparaffinized, formalin-fixed tissue

2007 ◽  
Vol 22 (4) ◽  
pp. 1228-1232 ◽  
Author(s):  
S. H. Nasr ◽  
G. S. Markowitz ◽  
A. M. Valeri ◽  
Z. Yu ◽  
L. Chen ◽  
...  
Keyword(s):  
Basement Membrane ◽  
Fixed Tissue ◽  
Thin Basement Membrane Nephropathy ◽  
Formalin Fixed Tissue ◽  
Formalin Fixed

Severe Antithrombin III Deficiency in an Infant Associated with Multiple Arterial and Venous Thromboses

1976 ◽  
Vol 36 (03) ◽  
pp. 495-502 ◽  
Author(s):  
Geoffrey Mendelsohn ◽  
Edward D. Gomperts ◽  
Dennis Gurwitz
Keyword(s):  
Antithrombin Iii ◽  
Adult Life ◽  
Rare Condition ◽  
Male Infant ◽  
Liver Cells ◽  
Liver Pathology ◽  
Fixed Tissue ◽  
Formalin Fixed Tissue ◽  
First Case ◽  
At Iii

SummaryInherited antithrombin III (AT-II, heparin cofactor) deficiency is a rare condition, presenting with thrombotic disease in adult life. This paper reports an 8 months old South African Black male infant with multiple large vessel venous and arterial thromboses, and E. coli septicaemia. This was associated with an extremely low plasma AT-II level. Micronodular cirrhosis and intracytoplasmic hyaline globules in the liver cells were present. These globules were eosinophilic, and PAS-positive after diastase. They measured approximately 5 μ to 30 μ in diameter, occurred singly in the liver cells and were located mainly in the periportal areas. The histological findings in the liver are similar to those observed in α1-antitrypsin (AAT) deficiency in which the intracytoplasmic globules represent accumulation of altered AAT. Immunochemical studies carried out on formalin fixed tissue failed to detect cross reaction material with anti-α1 antitrypsin or anti-AT III antiserum. This is the first case report of AT-III deficiency presenting in infancy. It is also the first case associated with distinctive liver pathology.The available data presented are insufficient to distinguish between an inborn defect and acquired causes of the severely depressed AT-III plasma level and the distinctive liver pathology.

Influence of technology parameters of free-cutting steel continuous casting on the billet internal structure

2020 ◽  
Vol 76 (2) ◽  
pp. 139-142
Author(s):  
A. T. Kunakbaeva ◽  
A. M. Stolyarov ◽  
M. V. Potapova
Keyword(s):  
Regression Analysis ◽  
Continuous Casting ◽  
Internal Structure ◽  
Sulfur Content ◽  
Manganese Content ◽  
High Quality ◽  
Correlation And Regression Analysis ◽  
Cutting Steel ◽  
Continuously Cast

Free-cutting steel gains specific working properties thanks to the high content of sulfur and phosphorus. These elements, especially sulfur, have a rather high tendency to segregation. Therefore, segregation defects in free-cutting steel continuously cast billets can be significantly developed. The aim of the work was to study the influence of the chemical composition of freecutting steel and casting technological parameters on the quality of the macrostructure of continuously cast billets. A metallographic assessment of the internal structure of cast metal made of free-cutting steel and data processing by application of correlation and regression analysis were the research methods. The array of production data of 43 heats of free-cutting steel of grade A12 was studied. Steel casting on a five-strand radial type continuous casting machine was carried out by various methods of metal pouring from tundish into the molds. Metal of 19 heats was poured with an open stream, and 24 heats – by a closed stream through submerged nozzles with a vertical hole. High-quality billets had a cross-sectional size of 150×150 mm. The macrostructure of high-quality square billets made of free-cutting steel of A12 grade is characterized by the presence of central porosity, axial segregation and peripheral point contamination, the degree of development of which was in the range from 1.5 to 2.0 points, segregation cracks and strips – about 1.0 points. In the course of casting with an open stream, almost all of these defects are more developed comparing with the casting by a closed stream. As a result of correlation and regression analysis, linear dependences of the development degree of segregation cracks and strips both axial and angular on the sulfur content in steel and on the ratio of manganese content to sulfur content were established. The degree of these defects development increases with growing of sulfur content in steel of A12 grade. These defects had especially strong development when sulfur content in steel was of more than 0.10%. To improve the quality of cast metal, it is necessary to have the ratio of the manganese content to the sulfur content in the metal more than eight.

Experience of clinical testing of personal telemedicine system ‘Obereg’ for remote monitoring of patients in intensive care units

2020 ◽  
pp. 52-58 ◽  
Author(s):  
A. A. Eryomenko ◽  
N. V. Rostunova ◽  
S. A. Budagyan ◽  
V. V. Stets
Keyword(s):  
Intensive Care ◽  
Intensive Care Units ◽  
Remote Monitoring ◽  
Clinical Testing ◽  
Transportation Of Patients ◽  
High Quality ◽  
Telemedicine System ◽  
Accuracy Of Measurements ◽  
Hospital Equipment

The experience of clinical testing of the personal telemedicine system ‘Obereg’ for remote monitoring of patients at the intensive care units of leading Russian clinics is described. The high quality of communication with the remote receiving devices of doctors, the accuracy of measurements, resistance to interference from various hospital equipment and the absence of its own impact on such equipment were confirmed. There are significant advantages compared to stationary patient monitors, in particular, for intra and out-of-hospital transportation of patients.

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