Histology of PLGA - (May/19/2007 )
Hi all,
I'm currently working on tissue engineering. I'm using PLGA as 3D model for cartilage regeneration, workin with human arthicular chondrocytes. But I have a problem, after seeding and culture the polymer for X days, to visualize the distribution and asses the new matrix formation, Hematoxilin-Eosin and Safranin O is used to asses that. But when I cut the polymer with microtomy for histology, it broke its intern structure and you cannot see the whole polymer, you only see the fragmented polymer. Here there is a visual example: http://www.fotozone.es/ficheros/2007/20/fotozone_26685.JPG
In papers, everybody used the same protocol for histology: fixation with formalin 4-10%, dehydratation with alcohols, xylenne, paraffin, and staining. I do what is suppose to do, but my question is if anybody knows how exactly is this proctol to obtain non-fragmented images from the polymer seeded with cell. Thanks in advance.
I'm currently working on tissue engineering. I'm using PLGA as 3D model for cartilage regeneration, workin with human arthicular chondrocytes. But I have a problem, after seeding and culture the polymer for X days, to visualize the distribution and asses the new matrix formation, Hematoxilin-Eosin and Safranin O is used to asses that. But when I cut the polymer with microtomy for histology, it broke its intern structure and you cannot see the whole polymer, you only see the fragmented polymer. Here there is a visual example: http://www.fotozone.es/ficheros/2007/20/fotozone_26685.JPG
In papers, everybody used the same protocol for histology: fixation with formalin 4-10%, dehydratation with alcohols, xylenne, paraffin, and staining. I do what is suppose to do, but my question is if anybody knows how exactly is this proctol to obtain non-fragmented images from the polymer seeded with cell. Thanks in advance.
Hi, in our lab, we fix the sample using cryostat medium. there are some cryostat medium available in the market. And, did you operate the cutting in room temperature ? In our lab, we did it in low temperature (-20oC). maybe you can try to use lower temperature when you cut your sample. hope it works
Thank you, but I already tried with cryostat medium "tissuetek" and cut by cryostat at -20ยบ, but I got the same results as with paraffin.
I'm currently working on tissue engineering. I'm using PLGA as 3D model for cartilage regeneration, workin with human arthicular chondrocytes. But I have a problem, after seeding and culture the polymer for X days, to visualize the distribution and asses the new matrix formation, Hematoxilin-Eosin and Safranin O is used to asses that. But when I cut the polymer with microtomy for histology, it broke its intern structure and you cannot see the whole polymer, you only see the fragmented polymer. Here there is a visual example: http://www.fotozone.es/ficheros/2007/20/fotozone_26685.JPG
In papers, everybody used the same protocol for histology: fixation with formalin 4-10%, dehydratation with alcohols, xylenne, paraffin, and staining. I do what is suppose to do, but my question is if anybody knows how exactly is this proctol to obtain non-fragmented images from the polymer seeded with cell. Thanks in advance.
Hi, in our lab, we fix the sample using cryostat medium. there are some cryostat medium available in the market. And, did you operate the cutting in room temperature ? In our lab, we did it in low temperature (-20oC). maybe you can try to use lower temperature when you cut your sample. hope it works