Thermo Scientific Varioskan Flash eng

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Varioskan Flash - Basic Operation Guide

Overview

The Varioskan Flash is a multimode microplate reader for various detection methods (absorbance, fluorescence, luminescence, time-resolved fluorescence). This guide provides a basic overview - the manual remains essential for detailed protocols!

Preparation

1. Power on the instrument

• Activate main power switch on device
• Wait for initialization to complete (approx. 2-3 minutes)
• Start "SkanIt" software on computer

2. Check basic settings

• Instrument should be warmed up (at least 30 min before measurement)
• Check temperature if temperature-controlled measurement is planned
• Verify lamp status (Xenon/Halogen depending on application)

Sample Preparation

Microplates

• Compatible plates: 96/384-well plates
• Plate type considerations:
  • Clear plates for absorbance
  • Black plates for fluorescence/luminescence
  • White plates for luminescence (optimal)

Sample Volume

• Minimum: ~50 μl (96-well), ~10 μl (384-well)
• Optimal: 100-200 μl (96-well), 20-50 μl (384-well)
• Avoid air bubbles!

Software Operation (SkanIt)

1. Create new protocol

• Select "New Protocol"
• Choose measurement type:
  • Absorbance: for ELISA, protein assays, cytotoxicity
  • Fluorescence: for Live/Dead assays, calcium imaging
  • Luminescence: for ATP assays, reporter genes

2. Set parameters

Absorbance:

• Select wavelength(s) (e.g., 450 nm for ELISA)
• Reference wavelength optional (e.g., 620 nm)

Fluorescence:

• Excitation wavelength (Ex)
• Emission wavelength (Em)
• Adjust gain/amplification

Luminescence:

• Set integration time
• Delay time if necessary

3. Define plate layout

• Mark wells: samples, standards, blanks, controls
• Dilution series can be calculated automatically
• Define replicates

Performing Measurements

1. Insert plate

• Carefully insert plate into carrier
• Note position A1 (usually top left)
• Close lid

2. Start measurement

• Click "Start" in software
• Do not open instrument during measurement!
• Measurement time depending on protocol: 30 seconds to several minutes

3. Kinetic measurements

• For time-dependent measurements (e.g., enzyme assays)
• Set interval and total duration
• Maintain constant temperature

Data Analysis

1. Check raw data

• Heatmap view for quick quality control
• Identify outliers
• Check blank values

2. Calculations

• Blank subtraction
• Create standard curve (for quantitative assays)
• Concentration calculations

3. Export

• Export data as Excel file
• Save graphs as images
• Save protocol for reproducibility

Important Tips

Before measurement

• ✅ Bring plate to room temperature
• ✅ Avoid condensation
• ✅ Minimize pipetting errors
• ✅ Include positive/negative controls

During measurement

• ❌ Do not open instrument
• ❌ Avoid vibration
• ❌ Avoid direct lighting (for fluorescence)

After measurement

• ✅ Save data immediately
• ✅ Document protocol
• ✅ If problems occur: repeat measurement

Common Applications

Assay Type	Mode	Typical Wavelengths	Plate Type
ELISA	Absorbance	450 nm (+ 620 nm Ref)	Clear
MTT/XTT	Absorbance	570 nm (+ 630 nm Ref)	Clear
FITC/GFP	Fluorescence	Ex: 485 nm, Em: 520 nm	Black
Cy5	Fluorescence	Ex: 640 nm, Em: 680 nm	Black
ATP Assay	Luminescence	-	White

Maintenance & Safety

Routine Care

• Keep instrument clean
• Clean plate carrier regularly
• Monitor filter/lamp status

Safety

• ⚠️ UV light: Protective eyewear for UV applications
• ⚠️ Note laser class
• ⚠️ Electrical safety: No water on instrument

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⚠️ IMPORTANT: This guide does NOT replace the official manual! Always consult the manufacturer's manual for specific protocols, troubleshooting, and advanced functions.