scholarly journals Two further alkaline phosphatase staining methods for immunohistochemistry.

1984 ◽  
Vol 37 (9) ◽  
pp. 1078-1079 ◽  
Author(s):  
P Fritz ◽  
H V Tuczek ◽  
J G Saal ◽  
G Wegner
Cells ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 127
Author(s):  
Wendy N. Erber

I reflect on my experience working with David Y. Mason in the Leukaemia Research Laboratories in the Nuffield Department of Pathology at the University of Oxford in the early 1980s. This was soon after the first monoclonal antibodies had been produced, which led to an exciting and productive time in biological discovery and pathology diagnostics. A specific focus in the laboratory was the development of immunoenzymatic staining methods that would enable monoclonal antibodies to be applied in diagnostic practice. This paper describes the work that led to the performance of immuno-alkaline phosphatase staining on blood and bone marrow smears, the success of which changed leukaemia diagnosis.


1994 ◽  
Vol 42 (3) ◽  
pp. 289-295 ◽  
Author(s):  
C M van der Loos ◽  
A E Becker

We present a method for an epi-illumination immunohistochemical double staining approach. The method combines the use of an immuno-alkaline phosphatase technique and the immunogold-silver technique, visualized with epifluorescence and epi-polarization illumination, respectively. Out of six tested alkaline phosphatase activity-revealing methods, only the reaction product obtained with the Becton Dickinson CAS Red kit showed an intense red fluorescence with a rhodamine filter set and no signal with epi-polarization illumination. The silver precipitate did not exhibit any signal with the rhodamine filter set. This allows separate observation and photographic recording of two antigens in one tissue section, an objective that cannot be achieved with conventional immunoenzyme double staining methods. The double epi-illumination approach presented is compatible with different immunoenzyme double staining protocols.


2007 ◽  
Vol 330-332 ◽  
pp. 23-26 ◽  
Author(s):  
Nam Sik Oh ◽  
Yun Ho Na ◽  
S. W. Ji ◽  
S.W. Song ◽  
S.H. Oh ◽  
...  

The aim of this paper was the HA and β-TCP powers were synthesized by a new wetchemical method using eggshell and phosphoric acid. The biocompatibility of synthesized natural HA, HA/β-TCP(50:50) and β-TCP derived from eggshell was compared with those of as commercial chemical powder with mesenchymal stem cells derived from human bone marrow. Development of crystalline phases of the mixtures was studied as functions of mixing ratio and temperature using X-ray diffractometer. The morphological characteristics of the calcined eggshell and synthesized powders were examined by scaning electron microscopy. The in-vitro cytotoxicity and cell attachment of sintered disks were examined using human bone marrowderived multipotent stem cells(hBMSCs). Cell response was characterized by MTT assay , Alkaline phosphatase stain and RT-PCR analysis. Pure HA was synthesized in the mixing ratio of 1:1.1 wt% at 900°C for 1h. the crystallization of HA was started at 800°C in the 1:1.1 mixing ratio, ant the HA phase was continued up to the high temperatures. In the ratio of 1:1.3 and 1:1.5 wt%, β-TCP was effectively synthesized at 900°C. In the 1:1.5 ratio, β-TCP phase was detected at 700°C, and complete crystallized β-TCP was observed above 900°C. At the higher temperature than 1000°C, the β-TCP was gradually decreased and α-TCP was observed. The HA and β-TCP disk does not exert cytotoxic effect on the hBMSCs undergoing osteoblastic differentiation. In addition, the hBMSCs are adhered on the surface of synthesized natural HA and β-TCP disk as successfully as on the culture plate or as commercial chemical HA and β-TCP disk. The hBMSCs adhered on either synthesized natural HA, β-TCP or as commercial chemical HA, β-TCP disk displays undistinguishable actin arrangement and cellular phenotypes, indicating that synthesized natural HA, β-TCP does not disrupt normal cellular responses. Analysis of differentiation of the hBMSCs cultured on culture plate, synthesized natural HA, β-TCP and as commercial chemichal HA, β-TCP disk shows that three matrices are able to support osteoblastic differentiation of the hBMSCs as accessed by alkaline phosphatase staining.


2000 ◽  
Vol 70 (5) ◽  
pp. 1793 ◽  
Author(s):  
William R Brown ◽  
Dixon M Moody ◽  
David A Stump ◽  
David M Colonna ◽  
Douglas J Kilgus

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 1579-1579
Author(s):  
Chunkang Chang ◽  
Chengming Fei ◽  
Youshan Zhao ◽  
Juan Guo ◽  
Xiao Li

Abstract Background The pathogenesis of MDS has not been completely understood, and insufficiency of the hematopoietic microenvironment can be an important factor. MSCs and osteoblasts are key components of the hematopoietic microenvironment. Studying osteoblastic differentiation of MSCs quantitatively may help to understand the pathogenesis of MDS. Methods 38 patients with MDS and 15 normal donors were investigated in this study. Osteoblastic differentiation assays were performed in 16 MDS cases and 8 controls. The expression of osteogenic differentiation markers were measured by real-time PCR. Alkaline phosphatase staining was performed with Alkaline Phosphatase staining kit after 3,7,14 days of incubation. ALP activity was assessed at 3, 7, and 10 days after osteogenic differentiation. Mineralization analysis was performed at 7, 14 and 21 days of osteogenic induction. The areas of mineralization were measured by Image-Pro Plus 6.0 software. Results Both MDS-MSCs and normal cells displayed same fibroblast-like morphology and similar antigen expression. The expression level of RUNX2 was significantly decreased in MSCs from MDS, compaired with normal controls, especially in lower-risk MDS. After osteogenic induction, lower-risk MDS showed lower alkaline phosphatase activity, less intense alizarin red S staining, and lower gene expression of osteogenic differentiation markers, however, higher-risk MDS was normal. Conclusions We concluded that impaired osteogenic differentiation of MSCs was seen mainly in patients with lower-risk MDS. It may contribute to the ineffective hamatopoiesis of MDS. Disclosures: No relevant conflicts of interest to declare.


1995 ◽  
Vol 16 (2) ◽  
pp. 125-129 ◽  
Author(s):  
TAKASHI KURIHARA ◽  
JUNKO HAYASHI ◽  
TOHRU MATSUOKA ◽  
AKIO ITO

Author(s):  
P D Mayne ◽  
A Ying Foo ◽  
Rosalind Michelson ◽  
I Z Kovar ◽  
S B Rosalki

We report on a new alkaline phosphatase isoenzyme abnormality occurring as an incidental finding in a male infant aged 4 months. Isoenzyme electrophoresis on cellulose acetate showed a prominent, diffuse alkaline phosphatase staining band in the α1-globulin position together with a second band in the α2/β region and minor ‘trailing’ in the intermediate α2 region. Normal liver and bone alkaline phosphatase were absent and intestinal phosphatase was not detected. On acrylamide gel electrophoresis a marked origin band was detectable suggesting the presence of high molecular weight enzyme. In addition, a series of compact bands in the α2/β position was present cathodal to the usual liver and bone isoenzymes. Total alkaline phosphatase activity was marginally elevated and was heat labile, L-phenylalanine resistant and partially L-homoarginine and L-leucine sensitive.


2005 ◽  
Vol 42 (2) ◽  
pp. 161-165 ◽  
Author(s):  
A. Barger ◽  
R. Graca ◽  
K. Bailey ◽  
J. Messick ◽  
L.-P. de Lorimier ◽  
...  

Aspiration of lytic bone lesions is an excellent diagnostic test in the initial evaluation of primary bone neoplasia. However, cytologically, it can be difficult to differentiate osteosarcoma (OSA) from other bone neoplasms, including fibrosarcoma, chondrosarcoma, synovial cell sarcoma, and plasma cell myeloma. The purpose of this study is to determine the sensitivity and specificity of alkaline phosphatase (ALP) staining to differentiate OSA from other tumors that express vimentin by immunocytochemistry or immunohistochemistry. ALP is a hydrolytic enzyme present in multiple tissues including liver, kidney, intestine, placenta, and bone. Hypothetically, neoplasms actively producing bone should be specifically positive for ALP staining. Unstained, cytologic specimens were incubated for 8-10 minutes with nitroblue tetrazolium chloride/5-bromo-4-chloro-3-indolyl phosphate toluidine salt-phosphatase substrate. A positive reaction stains the membrane of the cells gray to black. Samples were counterstained with a Romanowsky's stain to determine whether the sample was of representative cellularity. A total of 61 vimentin-positive neoplasms have been evaluated and confirmed histopathologically. Tumors that expressed vimentin and were positive for ALP included 33 OSAs, one multilobular tumor of bone, one amelanotic melanoma, and one chondrosarcoma. Tumors that expressed vimentin and were negative for ALP included chondrosarcomas (three of four), multiple fibrosarcomas, and multiple synovial cell sarcomas. The sensitivity is 100%, and the specificity is 89%. In conclusion, ALP appears to be a highly sensitive and fairly specific marker in the diagnosis of OSA.


2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
Mark Luigi Fabian Capati ◽  
Ayako Nakazono ◽  
Kohei Yamamoto ◽  
Kouji Sugimoto ◽  
Kajiro Yanagiguchi ◽  
...  

Tilapia type I atelocollagen (TAC) is a strong candidate for clinical application as its biological scaffold due to a high degeneration temperature and biologically safe properties. The aim of this study was to confirm the biological effects of TACin vitroon osteoblastic cells, simulating its clinical application. The proliferation and differentiation of typical preosteoblasts, MC3T3-E1 cells, were investigated using a microarray analysis, staining assay for mineralization, and real-time PCR analysis of the expression of mineralization-related genes. The mRNA expression of 10 genes involved in proliferation and differentiation increased after 3-day culture on an TAC gel, with an average balanced score ratio exceeding 1.5 compared to the control. After two weeks of culture, all three experimental groups showed stronger alkaline phosphatase staining than after one week. The genes expression of alkaline phosphatase, osteocalcin, and bone sialoprotein increased under the experimental conditions. The gene expression of osteopontin did not increase, and no statistical differences were noted among the three experimental groups. The present and previous findings suggest that TAC is not only a suitable alternative to collagen products originating from mammals but also a novel biomaterial with cell differentiation ability for regenerative medicine.


Sign in / Sign up

Export Citation Format

Share Document