Journal: Clinical Epigenetics

Article Title: Epigenetic landscape of testis specific histone H2B variant and its influence on sperm function

doi: 10.1186/s13148-021-01088-4

Figure Lengend Snippet: Semen parameters and chromatin compaction status of fertile individuals enrolled for ChIP-seq

Article Snippet: Slides were incubated with Acid detergent solution, pH 1.2 containing 0.08 N HCl, 0.15 M NaCl, 0.5% Triton X100 for 30 min at RT followed by staining with 0.25 mg/ml CMA3 (Enzo, Farmingdale, NY, USA) prepared in 0.1 M Citric acid, pH 7 containing 0.2 M Na 2 HPO 4 and 0.025 M MgCl 2 (McIlvaine buffer) for 30 min at RT in dark conditions.

Techniques: Concentration Assay

Journal: Clinical Epigenetics

Article Title: Epigenetic landscape of testis specific histone H2B variant and its influence on sperm function

doi: 10.1186/s13148-021-01088-4

Figure Lengend Snippet: Semen characteristics and chromatin compaction status of fertile and infertile men

Article Snippet: Slides were incubated with Acid detergent solution, pH 1.2 containing 0.08 N HCl, 0.15 M NaCl, 0.5% Triton X100 for 30 min at RT followed by staining with 0.25 mg/ml CMA3 (Enzo, Farmingdale, NY, USA) prepared in 0.1 M Citric acid, pH 7 containing 0.2 M Na 2 HPO 4 and 0.025 M MgCl 2 (McIlvaine buffer) for 30 min at RT in dark conditions.

Techniques: Concentration Assay

Journal: Clinical Epigenetics

Article Title: Epigenetic landscape of testis specific histone H2B variant and its influence on sperm function

doi: 10.1186/s13148-021-01088-4

Figure Lengend Snippet: Chromatin condensation in sperm of fertile and infertile men: Histone status of sperm was assessed by Aniline blue staining. Representative picture showing darkly stained immature sperm nuclei and light stained mature nuclei in fertile and infertile semen sample ( a -left panel). Protamine status was assessed by Chromomycin A3 staining. Representative picture of sperm from fertile and infertile semen samples shows bright fluorescing sperm nuclei which are CMA3 positive and dull fluorescing which are CMA3 negative ( b -left panel). Dot plot shows percentage of aniline blue ( a -right panel) and CMA3 positive sperm ( b -right panel) in the fertile (n = 18) and infertile individuals, namely, men with Asthenozoospermia (n = 10), Oligoasthenozoospermia (n = 10) and Oligozoospermia (n = 10); p **** < 0.0001

Article Snippet: Slides were incubated with Acid detergent solution, pH 1.2 containing 0.08 N HCl, 0.15 M NaCl, 0.5% Triton X100 for 30 min at RT followed by staining with 0.25 mg/ml CMA3 (Enzo, Farmingdale, NY, USA) prepared in 0.1 M Citric acid, pH 7 containing 0.2 M Na 2 HPO 4 and 0.025 M MgCl 2 (McIlvaine buffer) for 30 min at RT in dark conditions.

Techniques: Staining

Journal: Clinical Epigenetics

Article Title: Epigenetic landscape of testis specific histone H2B variant and its influence on sperm function

doi: 10.1186/s13148-021-01088-4

Figure Lengend Snippet: Correlation analysis between sperm chromatin compaction, various semen parameters and transcript levels of TH2B associated genes. Heatmap showing Spearman correlation analysis between percent aniline blue positive- (%AB+), percent CMA3 positive (%CMA3+), sperm motility, -concentration,-viability and transcript levels of CREM , CDYL , PRKAG2 , CATSPERB , TSGA10 and TSSK1B . p * < 0.05, p ** < 0.01, p *** < 0.001 and p **** < 0.0001

Article Snippet: Slides were incubated with Acid detergent solution, pH 1.2 containing 0.08 N HCl, 0.15 M NaCl, 0.5% Triton X100 for 30 min at RT followed by staining with 0.25 mg/ml CMA3 (Enzo, Farmingdale, NY, USA) prepared in 0.1 M Citric acid, pH 7 containing 0.2 M Na 2 HPO 4 and 0.025 M MgCl 2 (McIlvaine buffer) for 30 min at RT in dark conditions.

Techniques: Concentration Assay

Journal: Biomedicines

Article Title: Analyzing the Differential Impact of Semen Preparation Methods on the Outcomes of Assisted Reproductive Techniques

doi: 10.3390/biomedicines11020467

Figure Lengend Snippet: Sperm concentration, volume, WBC/HPG, TNMS (total normal motile sperms), normal % (normal morphology), HOS % (hypo-osmotic swelling), SDF% (sperm DNA fragmentation) and CMA3% levels in the whole studied population.

Article Snippet: The slides were then kept in a dark room for 20 min and were then stained with 150 mL of CMA3 (Merck 230752-10MG and Sigma-Aldrich Co, LLC, St. Louis, MO, USA C2659-10MG) (0.25 mg/mL) in McIlvain buffer.

Techniques: Concentration Assay

Journal: Biomedicines

Article Title: Analyzing the Differential Impact of Semen Preparation Methods on the Outcomes of Assisted Reproductive Techniques

doi: 10.3390/biomedicines11020467

Figure Lengend Snippet: TNMS (total normal motile sperms), HOS % (hypo-osmotic swelling), SDF% (sperm DNA fragmentation) and CMA3+% levels in the whole studied population after preparation of patients with TZs.

Article Snippet: The slides were then kept in a dark room for 20 min and were then stained with 150 mL of CMA3 (Merck 230752-10MG and Sigma-Aldrich Co, LLC, St. Louis, MO, USA C2659-10MG) (0.25 mg/mL) in McIlvain buffer.

Techniques:

Journal: Biomedicines

Article Title: Analyzing the Differential Impact of Semen Preparation Methods on the Outcomes of Assisted Reproductive Techniques

doi: 10.3390/biomedicines11020467

Figure Lengend Snippet: Values of CMA3+ % ( a ) from neat semen samples and ( b ) measured after spermatozoa selection using DGC-SU, DGC, SU and DGC-MACS. Semen samples separated here by ≤20 and >20% SDF. ** p < 0.01, *** p < 0.001.

Article Snippet: The slides were then kept in a dark room for 20 min and were then stained with 150 mL of CMA3 (Merck 230752-10MG and Sigma-Aldrich Co, LLC, St. Louis, MO, USA C2659-10MG) (0.25 mg/mL) in McIlvain buffer.

Techniques: Selection

Journal: The Journal of Reproduction and Development

Article Title: Preclinical evaluation of a new cryopreservation container for a limited number of human spermatozoa

doi: 10.1262/jrd.2017-086

Figure Lengend Snippet: CMA3 staining of fresh or freeze-thawed murine spermatozoa. CMA3-positive murine spermatozoa appear bright green, while CMA3-negative spermatozoa appear bright yellow. Bar: 20 µm. Picture of fresh spermatozoa (A) and spermatozoa in the straw (B) and the new container (C). Staining was repeated three times in the three groups (n = 3). The yellow color indicates non-damaged sperm and the green color indicates sperm with damaged nuclei. Spermatozoa with green heads were defined as CMA3-positive (∆) and those with yellow heads were defined as CMA3-negative (▲).

Article Snippet: Thereafter, the suspension was smeared on glass slides (S9441, Matsunami Glass) and smears were treated for 20 min with 200 µl of CMA3 solution (0.25 mg/ml CMA3 in Mcllvaline buffer [040-33731; Wako Pure Chemical Industries, Osaka, Japan]).

Techniques: Staining

Journal: Comparative Cytogenetics

Article Title: Male meiosis, heterochromatin characterization and chromosomal location of rDNA in Microtomus lunifer (Berg, 1900) (Hemiptera: Reduviidae: Hammacerinae)

doi: 10.3897/compcytogen.v5i1.1143

Figure Lengend Snippet: Microtomus lunifer . a–b C-banding and c–f Fluorescent banding: c, e DAPI and d, f CMA3. a Leptotene-zygotene. Very small C-positive dots can be observed in the autosomal chromatin; sex chromosomes are C-positive b Metaphase I. No C-positive bands can be detected c–d Pachytene e–f Diakinesis. No DAPI ( c, e )and neither CMA3-positive bands ( d, f ) can be detected, except for a small CMA3 bright band in one of the terminal regions of the largest autosomal pair. Arrows: sex chromosomes. Black arrowheads: smallest autosomal pair. White arrowheads: positive dots/bands. Bar = 10 µm

Article Snippet: Fluorescent banding with AT-specific DAPI and GC-specific chromomycin A3 (CMA3; Fluka BioChemika) was carried out as follows: after removal from freezer, the slides were placed immediately into cold 70% ethanol for 2 min. Then, they were transferred through 80% and 100% ethanol, 30 sec each, and air-dried.

Techniques: