TY - JOUR
T1 - Development of a method to preliminarily embed tissue samples using low melting temperature fish gelatin before sectioning
T2 - A technical note
AU - Ushida, Kaori
AU - Asai, Naoya
AU - Uchiyama, Kozo
AU - Enomoto, Atsushi
AU - Takahashi, Masahide
N1 - Funding Information:
We gratefully thank Reika Hatano, Shoma Tsubota, and Kenji Kadomatsu (Nagoya University) for providing cultured neurospheres. We also thank Masato Asai (Institute for Developmental Research, Aichi Human Service Center) and Daisuke Kuga (Anjo Kosei Hospital) for providing the images of mouse intestinal tissues. This work was supported by a Grant-in-Aid for Encouragement of Scientists (25930006 to K.U.) commissioned by the Ministry of Education, Culture, Sports, Science and Technology of Japan.
Publisher Copyright:
© 2018 Japanese Society of Pathology and John Wiley & Sons Australia, Ltd
PY - 2018/4
Y1 - 2018/4
N2 - Embedding of tissue samples that maintains a desired orientation is critical for preparing sections suitable for diagnosis and study objectives. Methods to prepare tissue sections include: (i) paraffin embedding or snap-freezing followed by microtome or cryostat sectioning; and (ii) agarose embedding followed by cutting on a vibrating microslicer. Although these methods are useful for routine laboratory work, preparation of small and fragile tissues such as mouse organs, small human biopsy samples, and cultured floating spheres is difficult and requires special skills. In particular, tissue specimen orientation can be lost during embedding in molds and subsequent sectioning. Here, we developed a method using low melting temperature (LM) gelatin either alone or mixed with agarose to preliminarily embed collected tissues that are either prefixed or unfixed, followed by conventional fixation, paraffin embedding, freezing, and sectioning. The advantage of the method is that the LM gelatin and its mixture with agarose can be handled at room temperature but quickly hardens at 4°C, which allows embedding, trimming, and arranging of small and fragile tissues in a desired orientation and are compatible with traditional stainings. Thus, this method can have various laboratory applications and can be modified according to the needs of each laboratory.
AB - Embedding of tissue samples that maintains a desired orientation is critical for preparing sections suitable for diagnosis and study objectives. Methods to prepare tissue sections include: (i) paraffin embedding or snap-freezing followed by microtome or cryostat sectioning; and (ii) agarose embedding followed by cutting on a vibrating microslicer. Although these methods are useful for routine laboratory work, preparation of small and fragile tissues such as mouse organs, small human biopsy samples, and cultured floating spheres is difficult and requires special skills. In particular, tissue specimen orientation can be lost during embedding in molds and subsequent sectioning. Here, we developed a method using low melting temperature (LM) gelatin either alone or mixed with agarose to preliminarily embed collected tissues that are either prefixed or unfixed, followed by conventional fixation, paraffin embedding, freezing, and sectioning. The advantage of the method is that the LM gelatin and its mixture with agarose can be handled at room temperature but quickly hardens at 4°C, which allows embedding, trimming, and arranging of small and fragile tissues in a desired orientation and are compatible with traditional stainings. Thus, this method can have various laboratory applications and can be modified according to the needs of each laboratory.
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U2 - 10.1111/pin.12652
DO - 10.1111/pin.12652
M3 - Article
C2 - 29465759
AN - SCOPUS:85042215021
VL - 68
SP - 241
EP - 245
JO - Acta Pathologica Japonica
JF - Acta Pathologica Japonica
SN - 1320-5463
IS - 4
ER -