Robust and artifact-free mounting of tissue samples for atomic force microscopy.
Immobilization of tissue-samples for atomic for microscopy (AFM) is typically done using either semi-dry tissue or by gluing the tissue sample down, both of which can introduce artifacts. Here, we describe the design of a Soft- Clamping Immobilizing Retainer of Tissue (SCIRT) for consistent and nondestructive immobilization of tissues for AFM analysis. We compare the performance of our SCIRT method with glue-immobilization for two difficult to handle tissue types: human trabecular meshwork (HTM) and rabbit cornea (RC). Our results demonstrate that the SCIRT method has several advantages, including: (i) allowing for small sample sizes, (ii) enabling continuous hydration, (iii) eliminating contact with glue or associated solvents, (iv) permitting sample recovery following measurement, and (v) ease of use. In conclusion, the SCIRT method is a simple and effective means of immobilizing soft, hydrated tissue samples consistently and without artifacts.
Department of Surgical and Radiological Sciences, School of Veterinary Medicine, University of California, Davis, One Shields Ave, Davis, CA.
Joshua T
JT
Morgan
Department of Surgical and Radiological Sciences, School of Veterinary Medicine, University of California, Davis, One Shields Ave, Davis, CA.
Vijay Krishna
VK
Raghunathan
Department of Surgical and Radiological Sciences, School of Veterinary Medicine, University of California, Davis, One Shields Ave, Davis, CA.
Sara M
SM
Thomasy
Department of Surgical and Radiological Sciences, School of Veterinary Medicine, University of California, Davis, One Shields Ave, Davis, CA; Department of Ophthalmology and Vision Science, School of Medicine, University of California, Davis, One Shields Ave, Davis, CA.
Christopher J
CJ
Murphy
Department of Surgical and Radiological Sciences, School of Veterinary Medicine, University of California, Davis, One Shields Ave, Davis, CA.
Paul
P
Russell
D000818:Animals; D016477:Artifacts; D006801:Humans; D018625:Microscopy, Atomic Force; D011817:Rabbits; D013048:Specimen Handling / Q000379:methods*
atomic force microscopy; cornea; sample preparation; tissue mechanics; trabecular meshwork