fluorescent probe prosense 680  (ProSense Inc)

Journal: Journal of Clinical Medicine

Article Title: Skeletal Muscle Subpopulation Rearrangements upon Rhabdomyosarcoma Development through Single-Cell Mass Cytometry

doi: 10.3390/jcm10040823

Figure Lengend Snippet: Generation of eRMS tumors in LSL-Kras G12D/+ ;Tp53 Fl/Fl mice. ( A ) Experimental workflow. LSL-Kras G12D/+ ;Tp53 Fl/Fl conditional mice were injected intramuscularly with Ad-Cre in order to induce genetic rearrangements, the constitutive activation of Kras and the inactivation of Tp53, which lead to eRMS tumor masses in a spatially and temporally controlled manner. ( B ) LSL-Kras G12D/+ ;Tp53 Fl/Fl conditional mice were injected with Ad-Cre and Ctrl vector. Tumor growth was assessed at three, five and seven weeks after Ad-Cre infection through FMT performed by intravitreal administration of the fluorescent probe (ProSense 680). This probe was activated by tumor cathepsin proteases to yield a fluorescence signal (black: low cathepsin activity; white: high cathepsin activity). ( C ) Representative H&E staining on histological sections from Ctrl and Ad-Cre infected muscles at three, five and seven weeks after Ad infection. ( D , E ) Immunofluorescence staining on sections of Ad-Cre infected muscles at three, five and seven weeks after infection. Sections were stained with antibodies raised against Caveolin-3 ( D ) and α-SMA ( E ). Scale bars: 100 µm.

Article Snippet: The emerging tumors were imaged by injecting a fluorescent probe (ProSense 680) that was specifically activated by cathepsin proteases, a tumor-specific enzymatic activity [ ].

Techniques: Injection, Activation Assay, Plasmid Preparation, Infection, Fluorescence, Activity Assay, Staining, Muscles, Immunofluorescence

Journal: Gut

Article Title: Detection of pancreatic cancer tumours and precursor lesions by cathepsin E activity in mouse models

doi: 10.1136/gutjnl-2011-300544

Figure Lengend Snippet: Cathepsin E (Cath E) activity can be detected efficiently in human pancreatic cancer xenografts in mouse. (A) Representative in-vivo image of human pancreatic cancer primary patient tumour grafts in mice showing brightfield (left) and fluorescence (Cath E activity) (right) signal at tumour location (n=4). (B) Representative ex-vivo image of mouse orthotopic tumour formed from cells (MDA PATC-3) isolated from human pancreatic cancer tumour grafts (A) showing fluorescence signal from pancreas with tumour (bottom) compared with a pancreas without a tumour (top) as well as kidney, muscle, bone, small intestine and lung (n=3). (C) Representative H&E staining of primary patient tumour grafts from (A) showing the presence of human tumour cells surrounded by the stromal components that characterise human pancreatic tumour. (D) Representative H&E staining of orthotopic pancreas tumour from (B) which shows the similar characteristics to human pancreatic ductal adenocarcinoma. Representative images of immunohistochemical localisation of Cath E of normal pancreas (E) from pancreas imaged in (B) and tumour (F) from tumour imaged in (B). Scale bar as shown in μm.

Article Snippet: 31 Both studies used a commercially available fluorescence probe (Prosense 680) that is sensitive to cathepsins B/H/L/S and plasmin.

Techniques: Activity Assay, In Vivo, Fluorescence, Ex Vivo, Isolation, Staining, Immunohistochemical staining

Journal: Gut

Article Title: Detection of pancreatic cancer tumours and precursor lesions by cathepsin E activity in mouse models

doi: 10.1136/gutjnl-2011-300544

Figure Lengend Snippet: Cathepsin E (Cath E) activity can be detected efficiently in genetically engineered mouse model (GEMM) tissues. (A) Representative ex-vivo image of mouse normal pancreas with spleen (left) in which no Cath E-activatable imaging probe was injected, mouse normal pancreas with spleen (middle) after 48 h of Cath E-activatable imaging probe injection, and GEMM (cLGL-KRasG12V with Bac-Ela-CreER mice) human pancreatic ductal adenocarcinoma (PDAC) tumour with spleen (right) after 48 h of Cath E-activatable imaging probe injection showing fluorescence signal specific to Cath E activity from the tumour and (B) fluorescence quantification of ex-vivo normal pancreas and GEMM (pooled data from both GEMM used) PDAC tumours showing approximately 15-fold increase in fluorescence in the pancreas tumours when compared with normal pancreas (n=5) (*p=0.0002). (C) Representative ex-vivo image of lung metastasis from mice with PDAC (p53 conditional deletion/LSL-KrasG12D/ Pdx1-Cre mice) without probe injection (left) and mice injected with Cath E-activatable imaging probe (right) showing fluorescence signal in lung metastasis when Cath E-activatable imaging probe is added (n=2). (D) Representative H&E staining and (E) representative image of immunohistochemical localisation of Cath E from tumour imaged in (A). (F) Representative H&E staining and (G) representative image of immunohistochemical localisation of Cath E from lung metastasis imaged in (C). Scale bar as shown in μm.

Article Snippet: 31 Both studies used a commercially available fluorescence probe (Prosense 680) that is sensitive to cathepsins B/H/L/S and plasmin.

Techniques: Activity Assay, Ex Vivo, Imaging, Injection, Fluorescence, Staining, Immunohistochemical staining

Journal: Gut

Article Title: Detection of pancreatic cancer tumours and precursor lesions by cathepsin E activity in mouse models

doi: 10.1136/gutjnl-2011-300544

Figure Lengend Snippet: Cathepsin E (Cath E) activity can be detected efficiently in mouse pancreas with pancreatic intraepithelial neoplasia (PanIN) lesions. (A) Representative ex-vivo Cath E-activatable imaging probe image of pancreas from genetically engineered mouse models (GEMM) with normal pancreas and pancreas with PanIN lesions (cLGL-KRasG12V with Bac-Ela-CreER mice) and pancreatic ductal adenocarcinoma (p53 conditional deletion/LSL-KrasG12D/Pdx1-Cre mice) injected with Cath E-activatable imaging probe. (B, C) Representative H&E staining of transgenic animals with normal pancreas and pancreas with PanIN but no tumours from (A). (D) Representative image of immunohistochemical localisation of Cath E from pancreas with PanIN but no tumours from (A). (E) Fluorescence quantification of ex-vivo pancreas showing approximately threefold increase in fluorescence in the pancreas with PanIN lesions when compared with normal pancreas (n=4) (*p=0.0058), and a significant increase in tumour fluorescence signal compared with the fluorescence signal from pancreas containing PanIN lesions but no tumour. Scale bar as shown in μm.

Article Snippet: 31 Both studies used a commercially available fluorescence probe (Prosense 680) that is sensitive to cathepsins B/H/L/S and plasmin.

Techniques: Activity Assay, Ex Vivo, Imaging, Injection, Staining, Transgenic Assay, Immunohistochemical staining, Fluorescence