Dreissenid Veliger Detection and Enumeration Technology to Improve Reliability and Sample Processing Using a Continuous Imaging Particle Analyzer (FlowCAM®®) Victoria Victoria M. M. Kurtz Kurtz Fluid Fluid Imaging Imaging Technologies Technologies Denise Denise Hosler Hosler Bureau Bureau of of Reclamation Reclamation Agenda ¾Introduction to Fluid Imaging Technologies, Inc. ¾FlowCAM® Technology ¾Specifications & Features ¾Applications Fluid Imaging Technologies ¾ Founded – 1999 ¾ Maine, USA (BLOS) ¾ Flow Cytometer And Microscope (FlowCAM) ¾ 500+ FlowCAMs sold in over 42 countries and on every continent ¾ Product Development ¾ FlowCAM VS Series ¾ Submersible ¾ VeligerCAM Dual Camera ¾ FlowCAM PV Series ¾ FlowCAM ES FlowCAM Models FlowCAM Applications ¾ Aquatic Research ¾Aquatic Research ¾ ¾ Discreet Discreetor orContinuous ContinuousSampling Sampling ¾ ¾ In Inlab lab ¾ ¾ In Insitu situ ¾ Monitoring/Detection of ¾Monitoring/Detection ofDreissenids Dreissenids ¾ Culture Studies ¾Culture Studies ¾ Algae Technology ¾Algae Technology ¾ Zooplankton Identification ¾Zooplankton Identification ¾ Training and ¾Training andEducation Education FlowCAM ¾ Rapidly Analyzes Particles in Fluids by Recording their Digital Image 30 frames per second (Upwards of 600 Images/Sec) ¾ Collects Size, Shape, Count, Concentration, and Data for All Particles Imaged – 2 µm to 2 mm ¾ Uses Data From Images to Construct Graphs Like Particle Size Distributions ¾ Allows the User to Examine the Microscopic Image of Any Particle Analyzed ¾ Provides for Automated Classification The partnership between Fluid Imaging Technologies, Inc. and the Bureau of Reclamation The photos provided by both the color FlowCAM® the dual camera VeligerCAM are very useful in a variety of studies VeligerCAM Configuration 15 mL Loading Pipette 12.5 mL Syringe Pump Dual Cameras 4X Microscope Objective Birefringent Filter Dual Camera Images Camera system • Dual B/W cameras • Camera 1 = normal bright field images • Camera 2 = birefringent images • Paired Images FlowCAM vs. VeligerCAM Images There have been improvements in the camera images The veligers now appear much brighter and identification is more efficient Increased Recovery of Veligers Average Recovery Before Changes - 75.1% Flow Rate (mL/min) Syringe Size (mL) Microscope Count VeligerCAM Count Percent Recovery 1.125 12.5 182 137 75.3% 1.125 12.5 202 156 77.2% 1.125 12.5 48 38 79.2% 1.125 12.5 77 53 68.8% Average Recovery After Changes - 98.0% Flow Rate (mL/min) 0.875 0.875 0.875 Microscope Count 53 51 50 Total Recovery 53 51 47 Percent Recovery 100% 100% 94% Percent Glycerin 40% 40% 40% Decreased Flow Rate Increased Viscosity A standard protocol was written by Scott O’meara and Kevin Bloom Microscope vs. VeligerCAM Images (Gray Scale Light) (Cross Polarized) Microscope (Gray Scale Light) (Cross Polarized) VeligerCAM The VeligerCAM provides greater detail of the specimen compared with microscope images Dye Study Internal Tissue The ability to see the veligers expel their internal tissue gave us clues for better sample management Denise Hosler and Kevin Bloom Turbulence Research Control Treatment Reclamation testing turbulence (high power water jets) to reduce veliger settlement VeligerCAM images show damage related to turbulence exposure: • Cracked, broken, or holes in shells • Expelled internal organs/ gapping shells Josh Mortensen and Sherri Pucherelli pH Degradation pH 5- The veligers are not birefringent but the VeligerCAM still takes photos under normal gray scale light pH 8- The veligers are birefringent and pair, making it quick and easy to identify Further investigation is underway Jamie Carmon and Kevin Bloom UV Study Control- Under gray scale light, the veliger internal tissue is still in tact Treatment- Under gray scale, the dark internal tissue is not present Further investigation is underway Zooplankton Study An efficient way to gather zooplankton and build libraries for water bodies Jamie Carmon and Kevin Bloom Thank You!
© Copyright 2024