(E-173) Coreview Imaging On a Needle for Point of Care Breast Cancer Diagnosis

Introduction::

In Low and Middle-income countries (LMIC), there is a lack of rapid point-of-care diagnostic devices for breast cancer. This can prevent patients from receiving results for months after core needle biopsy (CNB) procedures, with some individuals never getting a diagnosis or treatment plan due to socioeconomic and transportation barriers [1]. The Human Photonics Laboratory has partnered with UC Davis to create a Coreview ION (imaging on a needle) device that is designed to be a portable, rapid point-of-care diagnostic system for implementation in Ghana. The Coreview team developed a system that images core needle biopsies, using fluorescence stains and Microscopy with UV Surface Excitation (MUSE) imaging developed at UC Davis [2]. In traditional histopathology, artifacts and structure or tissue alteration can occur during tissue procurement, staining, fixation, and processing procedures [3]. The Coreview ION system minimizes tissue damage by eliminating destructive steps and also reduces processing time. Overall, our goal is to implement the Coreview Imaging on Needle (CVION) device for clinical use, providing high reliability and robustness. In taking steps towards this goal, we are streamlining our tissue processing procedures through the implementation of an automated liquid pump. The hydraulics are designed to replace manual tissue staining and washing procedures and once finalized, will reduce tissue preparation time and with future automation make the process turn-key. We also tested how tissue compression affects histopathologic interpretation.

Materials and Methods::

Materials: Hoechst (5 mg/mL) and Rhodamine B (5 mg/mL) stain and PBS (phosphate-buffered saline) solution. 10% Formalin Solution. Fresh pig breast tissues and FVB/N/Tg (TgMMTV-neu) mouse models. PLA plastic filament and Prusa Mini+ 3D printer. Plastic tubing (ID: 3.6 mm, OD: 6.3mm) and fluid control equipment (luer locks, check valves, etc.). Ximea xiD Series Camera and custom-made aluminum microscope fixture. 

Coreview Fixture: Frame consists of structured carbon polycarbonate plates attached to a custom microscope holder. The microscope is capable of 4x and 10x objective lens imaging under UV fluorescence with multi-axis movement control for the needle biopsy and biopsy compression.

Pump Design: Custom-made 3-D printed peristaltic pump. Controls are coded in C++ onto an Arduino. The pump works by rotating a hard ‘pusher’ to put pressure on pump channels, changing the volume of channels and displacing liquid. Liquid is expelled at the needle end. Our goal: effectively pump one droplet on command.

 

Staining Process:

1. 
   

   Dribble biopsy on the needle with 200 l of PBS

   
   


2. 
   

   Dribble 50 l of MUSE dye (Hoechst and Rhodamine mixture), wait 30 seconds

   
   


3. 
   

   Rinse biopsy with 200 l of PBS.

   
   

Pump Tests: 

1. 
   

   Select pump settings

   
   


2. 
   

   Fill pump tanks with chosen liquid

   
   


3. 
   

   Initialize mass of liquid containers

   
   


4. 
   

   Activate the pump.

   
   


5. 
   

   Measure mass of liquid pumped 

   
   

Compression Tests: 

1. 
   

   Acquire tissue with 14-gauge needle

   
   


2. 
   

   Wash and stain tissue

   
   


3. 
   

   Image tissue before compression

   
   


4. 
   

   Compress and image tissue at 50, 60, and 70% on needle

   
   


5. 
   

   Send tissues for review by Dr. Dintzis

   
   

 

Results, Conclusions, and Discussions::

Using pig breast tissue, our lab compressed samples to 50, 40 and 30 percent of the original thickness prior to device testing. With a MUSE imaging microscope, we photographed the biopsies at the various levels of compression and reviewed images with pathologist, Dr. Suzanne Dintzis. Tissues underwent conventional histopathology post-compression, the histopathologic diagnosis was not compromised by any of the three levels of biopsy compression. 

Our pump design consists of two liquid channels - one for staining and one for rinsing. We tested each at different volumetric and speed settings to determine how the speed at which liquid is pumped affects mass of liquid expelled repeatability. These tests were repeated across multiple liquids to identify potential failure modes under differing conditions. The data in figure 3.A shows that at higher speeds the pump has lower output than its intended volumetric setting. For the same channel and same speed setting, pumping PBS had less flow over time than distilled water.

 

Discussions

Further redesign is needed for our pump. The decreased repeatability of volume expelled at higher speeds could be due to back draft of liquid from high-speed volume displacements. The decreased efficiency concerning PBS needs addressing; we hypothesize that salt crystals in the liquid built up in the channels over time are blocking the flow of liquid [4]. Pump output variability leads to uneven staining and rinsing of biopsies, decreasing quality. With further testing, we wish to establish baseline repeatability across all speed and volume settings

We have demonstrated the first panoramic MUSE image from a fresh CNB using CoreView ION for making a tissue diagnostic image at the point of care. Ideally, the Coreview ION device will deliver results in less than three minutes after biopsy. We determine that tissue compression does not cause a noticeable decrease in quality of conventional histopathology using normal pig tissue.

Acknowledgements (Optional): : Special thanks to Drs. Fereidouni and Levenson at UC Davis pioneering MUSE imaging and assisting us in developing the Coreview system. Further thanks to the NIH NCI for their financial support through the R21 and R33 grans, our work would not have been possible without their help.

References (Optional): :

[1] P. Ssentongo et al., “Breast Cancer Survival in Eastern Region of Ghana,” Frontiers in Public Health, vol. 10, Jun. 2022, doi: https://doi.org/10.3389/fpubh.2022.880789.

[2] F. Fereidouni et al., “Microscopy with Ultraviolet Surface Excitation for Rapid Slide-Free Histology,” Nature Biomedical Engineering, 957-966 Dec. 2017, doi: https://doi.org/10.1038/s41551-017-0165-y

[3] V. Rastogi, “Artefacts: A Diagnostic Dilemma – A Review,” JOURNAL OF CLINICAL AND DIAGNOSTIC RESEARCH, 2013, doi: https://doi.org/10.7860/jcdr/2013/6170.3541.

[4] C. Tang and Y. Kim, “Optimization for a Centrifugal Pump Transporting Brine Using Orthogonal Design Based on CFD-DEM Simulation,” IOP Conference, 2021. doi:10.1088/1757-899X/1043/2/022061