PMID- 37578748
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20230816
IS  - 2076-2615 (Print)
IS  - 2076-2615 (Electronic)
IS  - 2076-2615 (Linking)
VI  - 13
IP  - 11
DP  - 2023 Jun 4
TI  - Validation of Calcein Violet as a New Marker of Semen Membrane Integrity in 
      Domestic Animals.
LID - 10.3390/ani13111874 [doi]
LID - 1874
AB  - Many fluorochromes routinely used in semen quality analysis emit in the green and 
      red channels, limiting their possible combination for multiple parameter 
      analysis. The use of fluorophores emitting in different light channels broadens 
      the possibilities of combination to expand the range of simultaneously evaluated 
      criteria. This is of great interest in cases of small ejaculated volumes, such as 
      those naturally occurring in roosters, small dog breeds and drones (Apis 
      mellifera). The purpose of this experiment is to establish Calcein Violet (CaV), 
      a blue fluorochrome, as a marker of viability and acrosomal integrity in domestic 
      animals in order to free the red and green channels. SYBR((R))14/Propidium Iodide 
      (PI) was used as reference dye, heat-treated samples as negative controls, serial 
      staining combination for validation and epifluorescence microscopy for 
      observation. Dead spermatozoa marked in red with PI showed no blue fluorescence 
      either from the head or the tail. Live spermatozoa showed a decreasing blue 
      emission from head to tail when single stained with CaV. Unreacted acrosomes 
      showed intense blue fluorescence irrespective of plasma membrane integrity. This 
      needs to be further confirmed for species with small and difficult to observe 
      heads. Establishment of CaV as a marker of membrane integrity by fluorescence 
      microscopy is a decisive first step towards further technical development and use 
      with flow cytometry.
FAU - Egyptien, Sophie
AU  - Egyptien S
AUID- ORCID: 0000-0001-8706-1049
AD  - Fundamental and Applied Research for Animals and Health Research Unit (FARAH), 
      Comparative Veterinary Medicine, Faculty of Veterinary Medicine, University of 
      Liege, Avenue de Cureghem 1, 4000 Liege, Belgium.
FAU - Dewals, Benjamin
AU  - Dewals B
AUID- ORCID: 0000-0003-1497-0799
AD  - FARAH, Veterinary Public Health, Faculty of Veterinary Medicine, University of 
      Liege, Avenue de Cureghem 1, 4000 Liege, Belgium.
FAU - Ectors, Fabien
AU  - Ectors F
AUID- ORCID: 0000-0003-2846-7019
AD  - FARAH, Veterinary Public Health, Faculty of Veterinary Medicine, University of 
      Liege, Avenue de Cureghem 1, 4000 Liege, Belgium.
FAU - Brutinel, Flore
AU  - Brutinel F
AUID- ORCID: 0009-0003-9958-149X
AD  - Fundamental and Applied Research for Animals and Health Research Unit (FARAH), 
      Comparative Veterinary Medicine, Faculty of Veterinary Medicine, University of 
      Liege, Avenue de Cureghem 1, 4000 Liege, Belgium.
FAU - Ponthier, Jerome
AU  - Ponthier J
AD  - Fundamental and Applied Research for Animals and Health Research Unit (FARAH), 
      Comparative Veterinary Medicine, Faculty of Veterinary Medicine, University of 
      Liege, Avenue de Cureghem 1, 4000 Liege, Belgium.
FAU - Deleuze, Stefan
AU  - Deleuze S
AUID- ORCID: 0000-0003-3076-5779
AD  - Fundamental and Applied Research for Animals and Health Research Unit (FARAH), 
      Comparative Veterinary Medicine, Faculty of Veterinary Medicine, University of 
      Liege, Avenue de Cureghem 1, 4000 Liege, Belgium.
LA  - eng
GR  - VETE-CCD/Fund for Scientific Research/
PT  - Journal Article
DEP - 20230604
PL  - Switzerland
TA  - Animals (Basel)
JT  - Animals : an open access journal from MDPI
JID - 101635614
PMC - PMC10252073
OTO - NOTNLM
OT  - acrosome
OT  - birds
OT  - epifluorescence
OT  - flow cytometry
OT  - insects
OT  - mammals
OT  - spermatozoa
OT  - viability
COIS- The authors declare no conflict of interest. The funders had no role in the 
      design of the study; in the collection, analyses, or interpretation of data; in 
      the writing of the manuscript; or in the decision to publish the results.
EDAT- 2023/08/14 12:42
MHDA- 2023/08/14 12:43
PMCR- 2023/06/04
CRDT- 2023/08/14 11:33
PHST- 2023/05/09 00:00 [received]
PHST- 2023/05/24 00:00 [revised]
PHST- 2023/06/02 00:00 [accepted]
PHST- 2023/08/14 12:43 [medline]
PHST- 2023/08/14 12:42 [pubmed]
PHST- 2023/08/14 11:33 [entrez]
PHST- 2023/06/04 00:00 [pmc-release]
AID - ani13111874 [pii]
AID - animals-13-01874 [pii]
AID - 10.3390/ani13111874 [doi]
PST - epublish
SO  - Animals (Basel). 2023 Jun 4;13(11):1874. doi: 10.3390/ani13111874.