title: An open source cryostage and software analysis method for detection of antifreeze activity.
abstract: The aim of this study is to provide the reader with a simple setup that can detect antifreeze proteins (AFP) by inhibition of ice recrystallisation in very small sample sizes. This includes an open source cryostage, a method for preparing and loading samples as well as a software analysis method. The entire setup was tested using hyperactive AFP from the cerambycid beetle, Rhagium mordax. Samples containing AFP were compared to buffer samples, and the results are visualised as crystal radius evolution over time and in absolute change over 30 min. Statistical analysis showed that samples containing AFP could reliably be told apart from controls after only two minutes of recrystallisation. The goal of providing a fast, cheap and easy method for detecting antifreeze proteins in solution was met, and further development of the system can be followed at https://github.com/pechano/cryostage.
authors.affiliation: Department of Biology, University of Southern Denmark, Campusvej 55, 5230 Odense M, Denmark. Electronic address: jloerup@gmail.com.
authors.forename: J L
authors.initials: JL
authors.lastname: Buch
authors.affiliation: Department of Science and Environment, Roskilde University, Universitetsvej 1, 4000 Roskilde, Denmark. Electronic address: hr@ruc.dk.
authors.forename: H
authors.initials: H
authors.lastname: Ramløv
mesh_terms: D000818:Animals; D021301:Antifreeze Proteins / Q000032:analysis* / Q000737:chemistry; D001517:Coleoptera; D003460:Crystallization; D012984:Software; D012996:Solutions
chemical_list: D021301:Antifreeze Proteins; D012996:Solutions
keywords: Antifreeze protein; Cryostage; Detection; Microscopy; Recrystallisation