Immuno-laser capture microdissection of frozen prolactioma sections to prepare proteomic samples.
Laser capture microdissection (LCM) technology combined with immunohistochemistry (immuno-LCM) is a valuable tool to obtain specific target cell populations and therefore this technique enables more accurate proteomic profile. In this study, we optimized the regular immuno-LCM technique to isolate and stain pure prolactin cells from either normal human pituitary (n=6) or prolactioma (n=11). Compared with the routine procedure, more intense and specific staining could be obtained when sections were pretreated with 0.2% Triton X-100 for 4 min. Interestingly, longer pretreatment (0.2% Triton X-100 for 10 min) or higher concentration (2% Triton X-100 for 4 and 10 min) greatly impaired labeling intensity and cell shape. Further scanning electron microscope study revealed that the component extracted from the cell surface by Triton X-100 was lipid. Using the optimized immuno-LCM technique, more pure prolactin cells could be isolated and prepared for further proteomic analysis. Taken together, we reported an optimized immuno-LCM technique that could effectively dissect pure target cells in different type pituitary adenomas for further proteomics analysis.
Liu
Yingchao
Y
Shanghai Neurosurgical Center, Department of Neurosurgery, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai 200040, China.
Wu
Jinsong
J
Liu
Sixiu
S
Zhuang
Dongxiao
D
Wang
Yongfei
Y
Shou
Xuefei
X
Zhu
Jianhong
J
D002469:Cell Separation; D005260:Female; D005629:Frozen Sections; D006801:Humans; D007150:Immunohistochemistry; D052682:Lactotrophs / Q000166:cytology / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D007834:Lasers; D008297:Male; D042282:Microdissection / Q000379:methods*; D017830:Octoxynol / Q000494:pharmacology; D010911:Pituitary Neoplasms / Q000378:metabolism* / Q000473:pathology; D015175:Prolactinoma / Q000378:metabolism* / Q000473:pathology; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods*
D020543:Proteome; D017830:Octoxynol