A simple method for double staining by immunofluorescence is described. If for double staining using monoclonal antibodies of the same species only one antibody is conjugated with FITC or TRITC, a combination of indirect and direct immunofluorescence is possible. For cell staining the following incubation steps are carried out: Monoclonal antibody I (unlabelled, mouse), anti-mouse immunoglobulin serum FITC- or TRITC-conjugated, normal mouse serum for blocking of free binding sites of the anti-mouse immunoglobulin, and monoclonal antibody II (mouse) which is conjugated with an alternative fluorochrome. The use of this method is demonstrated for investigation of single cell suspensions (performed as a slide test) and of cryostat sections.
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http://dx.doi.org/10.1016/S0065-1281(86)80052-6 | DOI Listing |
Nan Fang Yi Ke Da Xue Xue Bao
January 2025
Department of Rheumatology, Henan Provincial Hospital of Traditional Chinese Medicine// Second Affiliated Hospital of Henan University of Traditional Chinese Medicine, Zhengzhou 450002, China.
Objectives: To investigate the role of the BNIP3-PI3K/Akt signaling pathway in mediating the inhibitory effect of Decoction (BYHWT) on mitochondrial autophagy in human synovial fibroblasts from rheumatoid arthritis patients (FLS-RA) cultured under a hypoxic condition.
Methods: Forty normal Wistar rats were randomized into two groups (=20) for daily gavage of BYHWT or distilled water for 7 days to prepare BYHWT-medicated or control sera. FLS-RA were cultured in routine condition or exposed to hypoxia (10% O) for 24 h wigh subsequent treatment with IL-1β, followed by treatment with diluted BYHWT-medicated serum (5%, 10% and 20%) or control serum.
Clin Nephrol Case Stud
January 2025
Department of Medicine.
Minimal change disease (MCD) accounts for 10 - 15% of idiopathic nephrotic syndromes in adults. Chronic hepatitis C virus (HCV) infection is rarely ascribed as a cause of MCD and was previously associated with interferon-based therapy. MCD in treatment-naïve chronic HCV infection is extremely rare, with only 3 cases reported in the literature.
View Article and Find Full Text PDFClin Imaging
January 2025
Department of Radiology, Mayo Clinic, 200 First St SW, Rochester, MN 55905, United States of America.
Purpose: To evaluate henna as a durable skin marker on various skin tones for sonographic targeting and to identify the shortest duration of henna application needed for practical clinical workflow.
Materials And Methods: Prospective study applying seven henna lines through ultrasound (US) gel on the forearms of 15 healthy participants equally represented across the validated six-color bar tool. Color bar categories 1-2, 3-4, and 5-6 were designated low, moderate, and high-melanin groups, respectively.
Methods Appl Fluoresc
January 2025
Guangxi Medical University, No. 22, Shuangyong Road, Qingxiu District, Nanning, Guangxi, Nanning, Guangxi, 530021, CHINA.
Cell viability assessment plays a crucial role in biological research, pharmaceutical development, and toxicological identification. Here, we used GelRed, a sensitive and safer nucleic acid dye, to selectively label dead cells with red fluorescence (FL) thus distinguishing dead cells from live ones. Further more, the combined use of GelRed and SYTO 9 (another nucleic acid dye) enabled the clear differentiation in FL spectra between the two physiological statuses.
View Article and Find Full Text PDFCornea
January 2025
Department of Ophthalmology, University of Cyprus Medical School, Nicosia, Cyprus.
Purpose: To assess the impact of autologous serum (AS) tears at a 50% concentration on the ocular surface of patients with refractory dry eye disease (DED) because of Sjogren syndrome.
Methods: Twenty eyes of ten patients with severe immune-mediated DED were contralaterally randomized to receive either AS tears 50% or artificial tears between June 2021 and May 2023. Changes in tear stability, ocular surface staining, and in the morphology of the corneal sub-basal nerves were evaluated before treatment and at 1, 2, and 3 months after treatment using objective tests for DED and confocal microscopy.
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