EVOM3 Brochure

Quick Start Guide

EVOM3 Instruction Manual

STX2-PLUS Instruction Manual

Software for EVOM3 

Python digital output package

EVOM3 firmware upgrade package EVOM3 Rev 1.5 (current release)

FAQ

• Will the EVOM3 work with Endohm’s?

Yes, but the 99672 adaptor is required or the new EVOM3 cable 99916.

 

• Why would I want to use the blank function?

The blank feature is used when you want to subtract out any measurement that is not from the membrane, such as the electrode and fluid resistances.

 

• Does the EVOM3 system automatically calculate TEER?

No, TEER measurement requires an area calculation. To compute TEER, multiply the measured resistance by the appropriate surface area (below). For example, a 12 mm insert measures 565 Ω, the TEER is 565 Ω × 1.13 cm² = 638.5 Ω- cm². Here are the surface areas generally applicable to different transwell/insert formats: 6 well plate (24 mm inserts) 4.52 cm², 12 well plate (12 mm inserts) 1.13 cm², 24 well plate (6.5 mm inserts) 0.33 cm², 96 well plate (4.3 mm inserts) 0.14 cm².

 

• EVOM3 data is stored automatically when the last well is reached. How do I store the data when I only want to measure 8 of 96 wells?

Clear any data in memory by opening settings, store menu then press new plate, that will clear any prior readings. Return to the main screen, open the preview screen, select each well to measure (the selection turns green), place the electrode, then measure. When you’re done measuring the selected wells, open the settings, press the store screen menu, then press store new to save the plate data to the USB drive.

 

• How should you store the EVOM3 and electrodes if they will be exposed to UV light in a laminar hood for extended periods of time?

Take the EVOM3 out of the laminar hood after use. Next time, turn on the UV inside the hood. Once the hood is disinfected by UV, turn off UV, next spray 70-100% ethanol or isopropanol onto paper towel and wipe the EVOM3. Do not spray alcohol directly onto EVOM3.

 

• Why am I getting dashes as reading on EVOM3, even if I have the STX2-PLUS electrode inside the sample?

The electrode in the air or partially immersed in the liquid can show dashes since it records unstable read outs. The electrode tip portion (sensing region) must stay fully immersed. You may also notice unstable read outs when the electrode tip is not fully immersed. Make sure to select apical and basolateral volumes so that electrode tip stays fully immersed. You need to use apical and basolateral volumes greater than what is suggested by the insert manufacturer. For example, for Corning-24 well Transwell (example Corning 3470) we recommend minimum 300 µL on the top (apical) and 850 µL on the bottom (basolateral). [These volumes are a little more than the least required for STX2-PLUS electrode.]

Here are the steps:

Figure 1: STX2-PLUS Adjustment of Electrode Height. Rotate the front ring clockwise so that the electrode can enter to the maximum depth inside the well.

 

Figure 2: STX2-PLUS Electrode tip and liquid volume requirements. Make sure the electrode sensing tip (red boxed portions) on both blades stay fully immersed in a conductive liquid, such cell culture media or buffer during measurement. You need to have adequate apical and basolateral volumes to get a stable reading. Since STX2-PLUS stays hung, the increased volume must be used to make sure electrode sensing region fully immersed.

NOTE: You must use more liquid volumes than recommended by the insert manufacturer. Insert manufacturer’s recommended volumes will not keep electrode tip fully immersed.

[As mentioned as an example previously, for Corning-24 well Transwell (e.g., Corning 3470) we recommend using minimum 300 µL on top (apical) and 850 µL on bottom (basolateral). These volumes are a little more than the least required for STX2-PLUS electrode. You can check visually to make sure the apical and basolateral volumes are adequate to keep the electrode tips fully immersed, and then consistently use those volumes.]

Even if the unstable reading or dashes issue is still seen, the electrode most probably need chloriding. The chloriding refers to keeping electrode tips immersed in 3-6% sodium hypochlorite or bleach for 10-15 minutes followed by rinsing with distilled water. It is a part of STX2-PLUS maintenance and a critical maintenance process. Please refer to maintenance instruction below (step 1). **

 

• Can increasing or changing sample liquid volumes change my resistance values?

You can expect to see a change of raw resistance values. However, you subtract the blank values (blank Transwell with no cells) from the sample values (Transwell with cells). This way, you subtract the blank value with increased volume from samples with increased volume. Thus, any change of resistance contributed by increased volume is omitted. Consistently use the same volumes for all your samples in an experiment.

 

• Is there an electrode cleaning or maintenance instruction that I can follow?

Below are the steps that can be followed for STX2-PLUS cleaning or maintenance. Make sure you use enough liquid levels during cleaning or maintenance at least up to the red boxed region

1. Before using them, chloride the electrode by keeping the electrode tips immersed in 3-6% sodium hypochlorite (bleach) for 10-15 minutes. Chloriding needs to be done every 3 days when the electrodes are used frequently or after more than a week storage. **
2. Rinse with sterile DI water/buffer.
3. Optional step: Quick dip in 70% ethanol or isopropanol and quick dip in DI water/buffer.
4. Use the electrode for measurements.
5. Optional step in between measuring samples: Quick dip in 70% ethanol or isopropanol and quick dip in DI water/buffer.
6. After measurements soak/immerse electrode tips in 70% isopropanol or ethanol for 5-10 minutes.
7. Rinse with DI water. Let it air dry. Store electrode dry and in a place away from light/minimal light.
8. When used frequently, every week soak electrode tips in 1% Tergazyme for 15 minutes. Follow by rinse with DI water.
9. Next, chloride by keeping the electrode tips immersed in 3-6% sodium hypochlorite (bleach) for 10-15 minutes. (Same as step #1.)
10. Rinse with sterile DI water/buffer.
11. Use for measurements.
12. Repeat from step 5.

 

• Are there any other electrode handing instruction that WPI recommends?

	Do NOT hold the electrode by the cable. It can physically break the internal connections gradually.
	Hold the electrode by the arrowed region (plastic).
	Limit liquid immersion or liquid spray level somewhere up to here (maximum). You do not want the liquid to get inside and reach up to internal the cables or connectors that’s why. You can wipe with the rest of the electrode with a paper towel sprayed with isopropanol or ethanol (do not spray directly).

 

 

This unit conforms to the following specifications:

Type	Descriptions
Tissue Sampling Frequency	12.5 Hz
Sample Averaging	20 samples per second
Resistance Ranges	0 to 10,000 Ω  0 to 50,000 Ω  0 to 100,000 Ω +5%
Auto Mode	1 to 100,000 Ω auto current 2 μA, 4 μA, 10 μA
Resistance Resolution	0.1 Ω (under 200 Ω); 1 Ω (over 200 Ω)
Resistance Accuracy	0.1 Ω (under 200 Ω), 1 Ω (over 200 Ω) 0.1%  100,000 Ω ± 2 μA (to 105 KΩ)
Voltage Resolution	0.001 V, 0.1 mV
Accuracy Resistance	0.1 Ω (200 Ω); 1 Ω (above 200 Ω)
Accuracy voltage	± 0.1 mV
Current Levels	10,000 Ω ±10 μA 50,000 Ω ± 4 μA 100,000 Ω ± 2 μA Auto mode 1 to 100,000 Ω auto current 2 μA, 4 μA, 10 μA
Display Update Rate	0.5 seconds
Battery	3.7V Li-ion 2500 mAh**
Charging Period	5.5 hours (power off); 6 hours (run time)
Charge Current	200 mA
Power Consumption	~250 mA
Certifications	CE

** mAH means milliamp hours.

 

Inserts and Plates Compatible with the STX2-PLUS Electrodes

Corning	Millipore	Material	Membrane Diameter (mm)	Growth Surface Area (cm²)	Membrane Pore Size (μm)
3470			6.5	0.33	0.4
3472	PITP01250		6.5	0.33	3.0
3413	PCF Insert		6.5	0.33	0.4
3415	PITP 01250 PCF Insert		6.5	0.33	3.0
3421			6.5	0.33	5.0
3422	PIEP 01250 PCF Insert		6.5	0.33	8.0
3495	PIHT12R48* PET Insert		6.5	0.33	0.4
	PIHA012 50	HA Insert	6.5	0.33	0.45
	PICM012 50	CM Insert	6.5	0.33	0.4
3496	PISP12R48* PET Insert		6.5	0.33	3.0
	PIRP12R48* PET Insert		6.5	0.33	1.0
	PIMP12R48* PET Insert		6.5	0.33	5.0
	PIEP12R48* PET Insert		6.5	0.33	8.0
	PIXP01250 PCF Insert		6.5	0.33	12
	PIHP01250				1.0
	PITT01250				3.0

* Tri-supports 

Nunc	Pore size (μm)	Culture area (cm²)
140620	0.4	0.47
140627	3.0	0.47
140629	8.0	0.47

 

ThinCertTM	Membrane material	Pore size [µm]	Pore density [cm-2]	Optical membrane properties	TC surface treatment/Sterile	Multiwell plates/ThinCertTM per box
662640	PET	0.4	1 x 108	translucent	+/+	2/48
662641	PET	0.4	2 x 106	transparent	+/+	2/48
662610	PET	1.0	2 x 106	transparent	+/+	2/48
662630	PET	3.0	0.6 x 106	transparent	+/+	2/48
662631	PET	3.0	2 x 106	translucent	+/+	2/48
662638	PET	8.0	0.15 x 106	translucent	+/+	2/48

 

Millicell	Pore size (μm)	Qty/pk
MCHT24H48	0.4	48
MCRP24H48	1.0	48
MCSP24H48	3.0	48
MCMP24H48	5.0	48
MCEP24H48	8.0	48

  

BD Falcon	Membrane material	Pore size [µm]	Pore density [cm-2]	Optical membrane properties	TC plate (#wells)
353095	PET	0.4	2.0 ± 0.2 x 106	transparent	24
353104	PET	1.0	1.6 ± 0.6 x 106	transparent	24
353096	PET	3.0	8 ± 2 x 105	transparent	24
353097	PET	8.0	6 ± 2 x 104	translucent	24
353495	PET	0.4HD	100 ± 10 x 106	translucent	24
353492	PET	3.0HD	2.0 ± 0.2 x 105	translucent	24

 

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Ross, B. N.,

7 Reasons to Love the New EVOM3 for TEER Measurement

 

Great New Features of the EVOM3

The EVOM3 TEER Measurement System enables researchers to carry out experiments more efficiently by improving the workflow, and increasing the stability and accuracy of readings over that of the EVOM2. If you prefer to read the details, see the article "Why Choose an EVOM3 over an EVOM2." 

EVOM3: What's New?

Compact and Lightweight - This is a comparison between the EVOM3 and the EVOM2. At less than 1 lb., the EVOM3 is lightweight and portable. It has a sleek design with a touchscreen interface.

 

Smart Data Display and Foot Switch Control - See how easy it is to setup and use the foot switch to collect data.

 

Data Storage on USB Flash Drive - Save data as a Microsoft Excel files on the USB flash drive with the touch of a button. The data file can be accessed on a computer by plugging in the flash drive to a USB port.  

 

Improved Electrode Design - Compare the STX2 electrode and the NEW STX2-PLUS electrode. The new electrode stands vertically on the well plate, ensuring stable and consistent readings.