HUANG Zhaoshuai,LIN Wu,CHEN Xiaoming.. Observation of the fluorescent signals in the organs of mice[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2018, 48(9): 680-683.
Abstract： Objective: To observe the fluorescent signals in the liver, spleen and lung of mice and the quenching effect of autofluorescence by copper sulfate. Methods: ①Cell experiments: RG2 (rat glioma cell line) cells were divided into the control and treatment groups. Following maturity and attachment, two groups were fixed by 4% PFA. The treatment group was treated with 1.0 mmol/L copper sulfate solution (ammonium acetate buffer) for 10 min while the control group was not treated by the solution mentioned above. After sealed with fluorescent gel, the fluorescent intensity of these two groups was observed. ②Animal experiments: C57/BL6 mice were divided randomly into normal group and tail vein injection group. In tail vein injection group, the mice were injected with 100 µL of RG2 cell suspension and were sacrificed on the fourth day. After cardiac perfusion, the lungs, livers, and spleens were collected for frozen sections. In normal group, after cardiac perfusion, the lungs, livers, and spleens of mice were collected directly. The fluorescent signals of each organ and the effect of copper sulfate solution were observed. Results: After the treatment of copper sulfate, the fluorescent signals of RG2 reduced. The fluorescence distribution in each organ in mice was different; there was almost no autofluorescence in the lung; no obvious autofluorescence in the liver; marked autofluorescence in the spleen, which could not be eliminated by the treatment of copper sulfate solution. In contrast, the green fluorescence signals in the lung and liver in the tail vein injection group could be observed obviously. Conclusion: Copper sulfate can reduce the fluorescent intensity of cells and remove the background signal of spleen to a certain extent; the lung and liver can serve as better organs for the observation of fluorescence signals.
[1] RATHNAKAR B, RAO B S S, PRABHU V, et al. Laser-induced autofluorescence-based objective evaluation of burn tissue repair in mice[J]. Lasers Med Sci, 2018, 33(4): 699-707.
[2] 林潇, 倪浩棋, 黄李洁, 等. 常用脑出血动物模型的研究进展[J]. 温州医科大学学报, 2017, 37(9): 698-703.
[3] VAN DER HEYDEN J C, BIRNIE E, MUL D, et al. Increased skin autofluorescence of children and adolescents with type 1 diabetes despite a well-controlled HbA1c: results from a cohort study[J]. BMC Endocr Disord, 2016, 16 (1): 49.
[4] ERBEN T, OSSIG R, NAIM H Y, et al. What to do with high autofluorescence background in pancreatic tissues–an efficient Sudan black B quenching method for specific immunofluorescence labelling[J]. Histopathology, 2016, 69(3): 406-422.
[5] CROCE A C, BOTTIROLI G. Autofluorescence spectroscopy and imaging: a tool for biomedical research and diagno-sis[J]. Eur J Histochem, 2014, 58(4): 2461.
[6] DE VELD D, WITJES M, STERENBORG H, et al. The status of in vivo autofluorescence spectroscopy and imaging for oral oncology[J]. Oral Oncol, 2005, 41(2): 117-131.
[7] GEORGAKOUDI I, JACOBSON B C, MÜLLER M G, et al.
NAD (P) H and collagen as in vivo quantitative fluorescent biomarkers of epithelial precancerous changes[J]. Cancer Res, 2002, 62(3): 682-687.
[8] PELICANO H, XU R H, DU M, et al. Mitochondrial respiration defects in cancer cells cause activation of Akt survival pathway through a redox-mediated mechanism[J]. J Cell Biol, 2006, 175(6): 913-923.
[9] PALMER S, LITVINOVA K, RAFAILOV E U, et al. Detection of urinary bladder cancer cells using redox ratio and double excitation wavelengths autofluorescence[J]. Biomed Opt Express, 2015, 6(3): 977-986.
[10] PROSST R L, WEISS J, HUPP L, et al. Fluorescence-guided minimally invasive parathyroidectomy: clinical experience with a novel intraoperative detection technique for parathyroid glands[J]. World J Surg, 2010, 34(9): 2217-2222.
[11] PARAS C, KELLER M, WHITE L, et al. Near-infrared autofluorescence for the detection of parathyroid glands[J]. J Biomed Opt, 2011, 16(6): 067012.
[12] LADURNER R, SOMMEREY S, ARABI N A, et al. Intraoperative near-infrared autofluorescence imaging of parathyroid glands[J]. Surg Endosc, 2017, 31(8): 3140-3145.
[13] HILLARY S L, GUILLERMET S, BROWN N J, et al. Use of methylene blue and near-infrared fluorescence in thyroid and parathyroid surgery [J]. Langenbecks Arch Surg , 2018, 403(1): 111-118.
[14] SUN Y, YU H, ZHENG D, et al. Sudan black B reduces autofluorescence in murine renal tissue[J]. Arch Pathol Lab Med, 2011, 135(10): 1335-1342.
[15] OLIVEIRA V, CARRARA R, SIMOES D, et al. Sudan Black B treatment reduces autofluorescence and improves resolution of in situ hybridization specific fluorescent signals of brain sections[J]. Histol Histopathol, 2010, 25(8): 1017-1024.
[16] KIERNAN J A. Notes and queries[J]. Biotech Histochem, 2011, 86(3): 207-208.
[17] 冯亚平, 丁有权, 齐建国. 成年大鼠脊髓厚片自发荧光特异性消除新方法[J]. 解剖学研究, 2016, 38(6): 504-507.
