Hello Damien , I have got one question regarding VVG stain in my exam and it was bit hard to answer. Question was what is the colour of muscle looks like if before differential step.! Option was yellow , red/ magenta , green. I would be really appreciate it you can help me answer this question. Thankyou
Hi Anshu, This seems like quite a tricky question since following application of Verhoeff's Hx (first step in the VVG stain) the majority of structures are stained a very dark, almost black color, however, the presence of iodine within the Verhoeff's Hx mixture, can cause certain types of muscle to appear yellow/brown. Given the list of options to choose from you would therefore need to select "yellow". Alternatively, you can decide based upon exclusion of other options. The Red/Magenta color will not be present until after applying the Van Gieson solution, in which case the collagen (not muscle) will be stained red. Moreover, there is no green dye that I'm aware of used in combination with Verhoeff's Hx (would be interesting to attempt a brief stain in light green, but I think the Van Gieson mixture of picric acid and acid fuchsin provides a better contrast with Verhoeff's Hx-stained elastin fibres. Have you considered becoming a Trainee Histologist member of my RUclips channel? This will provide access to my online course in histological techniques. Good luck with your exams. D
Hello Damien. Does Van Gieson stain work on frozen sections? My muscle samples are embedded in OCT and frozed at -80Cº. I'm using Bouin fixation 20 min room temperature and after that I make the Van gieson stain. However the sections are not clear at all and change their color. Thanks a lot
Hi Yolanda, Yes. We have done this before. Our method and resulting images can be accessed via this link eprints.qut.edu.au/129086/ In brief, our method is as follows. Freshly acquired tissue samples were immersed in molds containing OCT then snap frozen using dry ice and stored at -80 degrees C. Sections of 3 microns in thickness were cut using a cryostat, transferred to adhesive slides and stored at -20 degrees C. To stain, sections were brought to room temperature over 5 minutes, rinsed three times for 5 min each in deionised water, before staining for 1 min in Van Gieson solution (0.1% w/v acid fuchsin dissolved in a saturated aqueous solution of picric acid). The stained sections were then carefully blotted using filter paper, dehydrated by 2 brief rinses in 100% alcohol (2-3 quick dips in each), cleared using two changes of xylene (2-3 minutes each), before being mounted beneath glass coverslip using plastic mounting medium (e.g., Entellan or Depex). The blotting and brief dehydration steps are vital to avoid loss of either stain.
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Thank you
This is so clear
Glad it was helpful!
Hello Damien , I have got one question regarding VVG stain in my exam and it was bit hard to answer. Question was what is the colour of muscle looks like if before differential step.! Option was yellow , red/ magenta , green. I would be really appreciate it you can help me answer this question.
Thankyou
Hi Anshu,
This seems like quite a tricky question since following application of Verhoeff's Hx (first step in the VVG stain) the majority of structures are stained a very dark, almost black color, however, the presence of iodine within the Verhoeff's Hx mixture, can cause certain types of muscle to appear yellow/brown. Given the list of options to choose from you would therefore need to select "yellow".
Alternatively, you can decide based upon exclusion of other options. The Red/Magenta color will not be present until after applying the Van Gieson solution, in which case the collagen (not muscle) will be stained red. Moreover, there is no green dye that I'm aware of used in combination with Verhoeff's Hx (would be interesting to attempt a brief stain in light green, but I think the Van Gieson mixture of picric acid and acid fuchsin provides a better contrast with Verhoeff's Hx-stained elastin fibres.
Have you considered becoming a Trainee Histologist member of my RUclips channel? This will provide access to my online course in histological techniques.
Good luck with your exams.
D
Hello Damien. Does Van Gieson stain work on frozen sections? My muscle samples are embedded in OCT and frozed at -80Cº. I'm using Bouin fixation 20 min room temperature and after that I make the Van gieson stain. However the sections are not clear at all and change their color. Thanks a lot
Hi Yolanda, Yes. We have done this before. Our method and resulting images can be accessed via this link eprints.qut.edu.au/129086/ In brief, our method is as follows. Freshly acquired tissue samples were immersed in molds containing OCT then snap frozen using dry ice and stored at -80 degrees C. Sections of 3 microns in thickness were cut using a cryostat, transferred to adhesive slides and stored at -20 degrees C. To stain, sections were brought to room temperature over 5 minutes, rinsed three times for 5 min each in deionised water, before staining for 1 min in Van Gieson solution (0.1% w/v acid fuchsin dissolved in a saturated aqueous solution of picric acid). The stained sections were then carefully blotted using filter paper, dehydrated by 2 brief rinses in 100% alcohol (2-3 quick dips in each), cleared using two changes of xylene (2-3 minutes each), before being mounted beneath glass coverslip using plastic mounting medium (e.g., Entellan or Depex). The blotting and brief dehydration steps are vital to avoid loss of either stain.
@@damienharkin Thank you Damien!! We will try as you say, and I hope this time work well!! Thank you so much