Ventana dp 200

Our slides were subjected to digitization using a whole slide imaging scanner known as Roche Ventana DP 200 (Roche Diagnostics/Ventana Medical Systems in Tucson, AZ, USA), and they underwent evaluation by both a scientist and a pathologist. This evaluation was conducted using the modified Remmele–Stegner Index (IRS) scale. The final score, which falls within the range of 0 to 12, is calculated by multiplying two factors: the percentage of cells or areas displaying positive staining (ranging from 0 to 4) and the intensity of staining (ranging from 0 to 3). In the evaluation of RUVBL1, both nuclear and cytoplasmic staining were assessed. A positive outcome was assigned to cases exhibiting staining in both the nuclei and cytoplasm of cells. Conversely, a negative outcome indicated the absence of staining in both the nuclei and cytoplasm. In the evaluation of HIF-1α, emphasis was placed on nuclear staining. A positive outcome was ascribed to instances where staining manifested within the cell nuclei; conversely, a negative outcome denoted the lack of staining within the nuclei.

The morphological and immunohistochemical features were analyzed on formalin-fixed and paraffin-embedded tissue sections. Samples of Patient 1 (9 weeks after the [⁸⁹Zr]Zr-Df-IAB22M2C PET/MRI) and Patient 3 (6 weeks after [⁸⁹Zr]Zr-Df-IAB22M2C PET/MRI) were stained with hematoxylin and eosin (H&E). Additionally, CD8 immunohistochemistry was performed on an automated immunostainer (Ventana Medical Systems, Tucson, AZ) according to the manufacturer's protocol using a CD8 antibody (clone C8/144B; Dako, Glostrup, Denmark). Appropriate positive and negative controls were used to confirm the adequacy of the staining. All samples were scanned with the Ventana DP200 (Roche, Basel, Switzerland) and processed with the Image Viewer MFC Application. Final image preparation was performed with Adobe Photoshop CS6.

Our slides were digitized using a whole slide imaging scanner (Roche Ventana DP 200) and were evaluated by both an image scientist and a pathologist. This evaluation was conducted using the modified Remmele–Stegner Index (IRS) scale. The final score, ranging from 0 to 12, is derived from the multiplication of two factors: the percentage of cells/areas stained positively (ranging from 0 to 4) and the staining intensity (ranging from 0 to 3). The scans were subsequently saved in our internal repository. The Evaluate Cutpoints application was utilized to determine the cutpoints. The threshold values for high and low membranous expression of β-Catenin were as follows: <8 (abnormal); ≥8 (normal). The nuclear expression of UBB and UBC was analyzed based on the presence or absence of nuclear staining in cells. The threshold values for membranous expression of β-Catenin were as follows: <8 (abnormal); ≥8 (normal). In turn, a cutpoint of ≥1 was used to indicate the presence of nuclear expression of UBB and UBC (<1, absent). In dual-IHC staining, nuclear UBB expression and membrane expression of β-Catenin were assessed based on the presence and absence of staining to test the hypothesis of the present study.

After MALDI-MSI measurements, matrix was removed from tissues by washing in 100% methanol for 2 min. Tissues were stained with hematoxylin and eosin (HE) for pathological evaluation. Whole slide scanning was performed on HE sections for digital evaluation of the tissues using a Ventana DP 200 slide scanner (Roche Diagnostics International AG, Rotkreuz, Switzerland). Representative tumor and stroma regions were annotated on whole slide images using the software QuPath (Bankhead et al. 2017 (link)) and regions were exported for further analysis using the SCiLS Lab export function (Bruker Daltonik GmbH, Bremen, Germany).

The FSW device (PiezoWave) was purchased from Richard Wolf GmbH (Knittlingen, Germany). A 500 kHz × 64 mm focused piezoelectric transducer (H-104G) and its fundamental and third harmonic resonance impedance matching network were acquired from Sonic concepts, Inc. (Washington, United States). A RF Power Amplifier (40AD1) was obtained from Amplifier Research Inc. (Pennsylvania, United States). A RF multifunction power meter (4421) and its directional power sensor (4025) were procured from Bird Technologies Co. (Ohio, United States). A function/arbitrary waveform generator (33120A) was purchased from Agilent Technologies, Inc. (California, United States). An oscilloscope (LT354ML) was obtained from LeCroy Co. (New York, United States). An immersion planar ultrasound transducer (1 MHz, A392S-SU) and manually controlled ultrasound pulser-receivers (5072PR) were acquired from Olympus Co. (Tokyo, Japan). A slide scanner (Ventana Dp200) and its software (Ventana Image Viewer v3.2) were obtained from F. Hoffmann-La Roche Ltd. (Basel, Switzerland). A microplate reader (Infinite 2000 Pro) and its software (i-control) were procured from Tecan Austria GmbH Co. (Grodig, Austria).