The Attune®
Autosampler

Easy operation through multiple-sample processing

The Attune® Autosampler, an optional accessory to the Attune® Acoustic Focusing Cytometer, enables rapid processing of up to 384 samples.

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Learn about the Attune® Cytometer

Watch the Attune® Autosampler Video Demo

 The Attune® Autosampler enables multiple-sample processing and is compatible with many different formats including 96- and 384-well plates and deep-well plates. To transition between use of the autosampler and individual tubes by the flip of a switch. Its intelligent probe leads to minimum clogging, carryover, and prevents instrument damage.

Broad compatibility

Compatible with many different plate formats, including 96-well, 384-well, and deep-well plates


Intelligent Probe

Intelligent probe design minimizes clogging and carryover (<0.5%) and prevents damage to the instrument


Mixing by Aspiration

Mixing sample by aspiration instead of shakings ensures homogeneity of the sample and maintains cell viability
See why mixing sample by aspiration is better >

Download the Attune® Autosampler flyer

Auto cleaning

Performs automated cleaning when the instrument is shutting down


Consistent Data

Minimal variation regardless of sampling method (tube vs. plate) and collection rates


Free software

Analyze data on any computer from anywhere at any time—no limit on the number of users

Software

Setting up an experiment with Attune® software is simple. Using a virtual plate layout view, you can create compensation wells, define instrument settings, and identify samples. The software also allows you to define multiple experiments on a single plate and recover unused sample (for samples collected or acquired in tube mode only).

The intuitive software is easy to use and enables quick data analysis. For example, a heat-map view allows rapid screening and confirmation of samples while the instrument is acquiring data (Figures 2 and 3).

The Attune® software is free and can be downloaded online. Multiple users can access the software-no license fees or dongles are required.

  Number of washes and % carryover
Mode 123
Standard.01.01.01
High throughput.02.02.02

Table 1. Minimal carryover using the Attune® Autosampler. Jurkat cells at a concentration of 1 x 106 cells/mL were aliquoted into a 96-well, vbottom plate and sampled using the Attune® Autosampler. Samples were analyzed on the Attune® Acoustic Focusing Cytometer using a collection rate of Standard Mode (200 μL/min) and High Throughput (500 μL/min). Each sample was mixed once, and the Attune® Autosampler was washed 1–3 times prior to sampling the next well. Percent sample carryover was calculated.

 

Increasing the Number of Mixing Cycles Does Not Adversely Affect Cell Viability

 

Percent Dead  Cells (%)

Number of Mix Cycles

LWBNIH/3T3
0.7534.10
1.7832.83
2.7433.52
3.7432.75
4.7433.26
5.7531.58

Table 2. Gentle sample mixing using the Attune® Autosampler. Increasing the number of mixing cycles does not adversely affect cell viability. Ammonium chloride–lysed whole blood (LWB) and NIH/3T3 (live/heat-treated) cells were stained with 2 μg/mL propidium iodide, loaded in triplicate in a 96-well, v-bottom plate. Prior to acquisition samples were mixed 0–5 times by the Attune® Autosampler, and then samples were analyzed using Standard mode collection rates (100 µL/min for NIH/3T3, 200 µL/min for LWB) on the Attune® Acoustic Focusing Cytometer. Propidium iodide was excited using a 488 nm laser and fluorescence emission was collected using a 640 nm longpass filter. Minimal variation was observed in both samples, regardless of cell type and the number of mix cycles used prior to acquisition.

 

Consistent Results Regardless of Sampling Method

Figure 1. Consistent results regardless of sampling method. Whole blood lysed with ammonium chloride was labeled with mouse anti-human CD45 Pacific Orange™, mouse anti-human CD4 FITC and mouse anti-human CD8 RPE antibody conjugates. Labeled samples were analyzed on a blue/violet-configured Attune® Acoustic Focusing Cytometer equipped with a 488 nm laser for fluorescence excitation of FITC (530 BP) and RPE (574/24 BP) and a 405 nm laser for Pacific Orange™ dye (603/48 LP). Identical samples, including compensation controls, were analyzed using either (A) tube mode or (B) plate mode at a collection rate of Standard 200 μL/min. Lymphocytes were gated using a CD45 vs. side scatter plot and analyzed for expression of CD4 and CD8 antigens. Minimal variation was observed between analysis in a tube alone and on a plate running on the Attune® Autosampler.

 

Consistent Results, Well-to-Well

The Attune® Autosampler heat map function identifies variation within a parameter across a 96-well plate.

Figure 2. Consistent well-to-well results: the Attune® Autosampler heat map function identifies variation within a parameter across a 96-well plate. Live and heat-killed THP-1 cells were stained with 2 μg/mL propidium iodide, aliquoted into a 96-well, V-bottom plate, and run using a collection rate of Standard 500 μL/min with 2 mix cycles per well and 2 rinse cycles between wells. Propidium iodide was excited using a 488 nm laser (640 LP). The heat map image graphically represents the percent propidium iodine positive cells (dead cells) using the color gradient indicated in (A). Redcolored wells indicate 0% propidium iodide–positive cells within the sample analyzed from that well, whereas magenta-colored wells indicate a sample containing 100% propidium iodide–positive cells. The text overlaid on each well in the heat map (B) is the measured percentage of dead cells from each individual well within the plate. Minimal variation is observed in propidium iodide fluorescence across the entire plate, with a coefficient of variation equal to 1.44% for the entire data set (96 wells).

Physical Characteristics

  • Footprint (H x W x D): approximately 16 x 11 x 11 in (40 x 29 x 29 cm)
  • Weight: approximately 35lb (16 kg)
  • Operating temperature: 50-95°F (15 to 30°C)
  • Operating Humidity: <80% noncondensing
  • Electrical requirements: 100-240 VAC, 50/60 Hz, <300 W

 

Software/Computer Requirements:

  • Attune® Cytometric Software Version 2.0 or higher
  • Windows® 7 Operating System

 

Compatible Plate Types:

  • 96-deep-well (flat, round, and V-bottom)
  • 96-well standard depth (flat, round, and V-bottom)
  • 384-well standard depth (flat, round, and V-bottom)
  • 384-deep-well (flat, round, and V-bottom)

 

Processing Time:

  • <45 minutes for 96-well plate, using High Throughput mode
  • <60 minutes for 96-well plate, using standard mode, 2 wash cycles
  • <180 minutes for 384-well plate using High
    Throughput mode
  • <260 minutes for 384-well plate using Standard Mode, 2 wash cycles

Carry Over:

  • Less than 0.5%

 

Mixing Cycles:

  • Each well will be mixed via full aspiration (not shaking)

 

Wash Cycles:

  • User-defined number of wash cycles dependent upon plate-processing protocol and time to acquire plates

 

Minimum Sample Required:

  • 50 μL for 96-well plates

 

Minimum Dead Volume:

  • 30 μL

 

Fluidics Requirements:

  • On board fluidics tanks 800 mL total
  • Capable of running four 96-well plates in standard mode with two washes per well