Journal: Laboratory investigation; a journal of technical methods and pathology

Article Title: Reproducible, high-dimensional imaging in archival human tissue by Multiplexed Ion Beam Imaging by Time-of-Flight (MIBI-TOF)

doi: 10.1038/s41374-022-00778-8

Figure Lengend Snippet: MIBI-TOF staining panel.

Article Snippet: Individual metal labeled antibodies for IHC were also obtained from Ionpath: CD3 (D7A6E), CD8 (C8/144B), CD68 (D4B9C), Pax5 (D7H5X), PanCK (AE1/AE3).

Techniques: Staining

Journal: Current Oncology

Article Title: A Preliminary Analysis of Circulating Tumor Microemboli from Breast Cancer Patients during Follow-Up Visits

doi: 10.3390/curroncol31090421

Figure Lengend Snippet: Classification of clustered CTC (CTM) by image-based analysis. After immunofluorescent staining to label-specific markers including the PanCK, Hoechst, CD45, and CD3 on CTC/CTM, the cell-loaded SACA-chips were scanned by the automatic imaging platform (CytoSCM imaging system). Digitally labeled images of CTC/CTM in different channels (PanCK: FITC channel; Hoechst 33342: UV365 channel; CD45: PE channel, CD3: APC channel, and Merged channel) were further analyzed to detect, enumerate, and identify different CTC subpopulations. ( A ): Output images of different channels to show the individual CTC identified by the platform; ( B ): output images of different channels to show four subtypes of clustered CTC identified by the platform, including the CTC cluster with only PanCK+ CTC, the clustered CTC (PanCK+) accompanied by only CD45+ leukocytes in which the ratio of PanCK+ CTC and CD45+ leukocytes was less than 1, the small group of clustered CTC (PanCK+) accompanied by CD45+ leukocytes and CD3+ lymphocytes, in which the ratio of PanCK+ CTC and the sum of CD45+ leukocytes and CD3+ lymphocytes was 1 or more than 1; the large group of clustered CTC (PanCK+) accompanied by CD45+ leukocytes and CD3+ lymphocytes, in which the ratio of PanCK+ CTC and the sum of CD 45+ leukocytes and CD 3+ lymphocytes was less than 1. The red circle in ( A ) was displayed to show the identified area of the targeted CTC in different channels by the automatic imaging platform. The green circle in ( B ) was displayed to show the identified CTC in different channels by the automatic imaging platform. The orange circle in ( B ) was displayed to show the identified leukocytes in different channels by the automatic imaging platform. The red circle in ( B ) was displayed to show the identified lymphocytes in different channels by the automatic imaging platform.

Article Snippet: Resuspended cells in PBS were counted and stained by Hoechst 33342 (Thermo Fisher Scientific Taiwan Co., Ltd., Taipei, Taiwan), anti-pan cytokeratin (PanCK) conjugated with fluorescein isothiocyanate (FITC) (Catalog number: IR222-861-FITC, iReal Biotechnology Inc., Hsinchu, Taiwan), anti-CD45-phycoerythrin (PE) (Catalog number: 12-0459-42, Thermo Fisher Scientific Taiwan Co., Ltd., Taipei, Taiwan), anti-CD3-APC (Catalog number: E-AB-F1001E), anti-CD8-APC (Catalog number: E-AB-F1110E), and anti-estrogen receptor alpha (ER) -PE (Catalog number: AB209288, Abcam, Blossom Biotechnologies Inc., Taipei, Taiwan) in the dark at room temperature to label the nucleus, CTC/CTM, leucocytes, lymphocytes, and the estrogen receptor (ER) on CTC/CTM.

Techniques: Staining, Imaging, Labeling

Journal: Current Oncology

Article Title: A Preliminary Analysis of Circulating Tumor Microemboli from Breast Cancer Patients during Follow-Up Visits

doi: 10.3390/curroncol31090421

Figure Lengend Snippet: Expressions of the estrogen receptor and CD8 in the CTM identified in breast cancer patients. ( A ): Output images from four channels of our chip-based imaging system together with the merged and bright field images to illustrate the PanCK+ CTC, nuclei, ER expression (positive), and CD8 expression (positive) in the CTM identified from patient 4 who was diagnosed as disease-free (DF) during follow-up visits; ( B ): output images from four channels of our chip-based imaging system together with the merged and bright field images to illustrate the PanCK+ CTC, nuclei, ER expression (negative), and CD8 expression (negative) in the CTM identified in patient 5 who was also diagnosed as disease-free (DF) during follow-up visits; ( C ) output images from four channels of our chip-based imaging system together with the merged and bright field images to illustrate the PanCK+ CTC, nuclei, ER expression (negative), and CD8 expression (negative) in the CTM identified in patient 7 who was experiencing hormonal therapy and diagnosed with disease recurrence (DR) during follow-up visits. After immunofluorescent staining to label-specific markers including the PanCK, Hoechst, and CD45 to identify the CTM, samples were also examined for the expression of CD8 and ER on the identified CTM. The cell-loaded SACA-chip was scanned by the automatic imaging platform CytoSCM. The digitally labeled area of the identified target in images from four different channels (PanCK-FITC; Hoechst 33342-UV; ER-PE; CD8-APC), merged images, and bright field images are circled by the yellow line.

Article Snippet: Resuspended cells in PBS were counted and stained by Hoechst 33342 (Thermo Fisher Scientific Taiwan Co., Ltd., Taipei, Taiwan), anti-pan cytokeratin (PanCK) conjugated with fluorescein isothiocyanate (FITC) (Catalog number: IR222-861-FITC, iReal Biotechnology Inc., Hsinchu, Taiwan), anti-CD45-phycoerythrin (PE) (Catalog number: 12-0459-42, Thermo Fisher Scientific Taiwan Co., Ltd., Taipei, Taiwan), anti-CD3-APC (Catalog number: E-AB-F1001E), anti-CD8-APC (Catalog number: E-AB-F1110E), and anti-estrogen receptor alpha (ER) -PE (Catalog number: AB209288, Abcam, Blossom Biotechnologies Inc., Taipei, Taiwan) in the dark at room temperature to label the nucleus, CTC/CTM, leucocytes, lymphocytes, and the estrogen receptor (ER) on CTC/CTM.

Techniques: Imaging, Expressing, Staining, Labeling

Journal: Analytical Chemistry

Article Title: Lipidomic Phenotyping Of Human Small Intestinal Organoids Using Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Imaging

doi: 10.1021/acs.analchem.3c03543

Figure Lengend Snippet: Characterization of human SIOs using MALDI-MSI and (immuno)histochemical staining. (A) Characterization of crypt-like and villus-like human SIOs using MALDI-MSI different histological stainings. Brightfield images representing different phenotypes of human SIOs, including CL and VL organoids (scale bar: 500 μm). MALDI-MSI measurements were performed in negative ion mode, where several lipid species were visualized in both CL and VL SIO: (red) PE 36:2 [M – H] − ( m / z 742.5), (green) PI 38:4 [M – H] − ( m / z 885.5) and (blue) PI 36:1 [M – H] − ( m / z 863.6). Representative stainings for organoid morphology (H&E) and intestinal differentiation markers in CL- and VL-cultured SIO. Alkaline phosphatase staining of the brush border of enterocytes (blue) and Alcian blue-stained goblet cells or mucus (light blue). (B) MALDI-IHC of CL organoids using a PC-MT of PanCK. A consecutive H&E staining was performed on the same sample after measurement. A conservative IHC staining of PanCK (brown) was performed on a consecutive organoid section to validate the MALDI-IHC results (scale bar: 500 μm).

Article Snippet: Sections were stained with pan cytokeratin (panCK) PC-MT antibody probe (Ambergen Inc., Boston, MA) according to the manufacturer’s instructions.

Techniques: Staining, Cell Culture, Immunohistochemistry