Dreissenid Veliger Detection and Enumeration Technology to Improve Reliability and Sample Processing

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!