96-Well Plate

What is a 96-well plate?

A 96-well plate is a rigid tray moulded with 96 small wells in eight rows and twelve columns. Each well behaves like an independent miniature test tube, so one plate can hold 96 separate samples, reactions or conditions that are pipetted, incubated and read together. It is the single most common vessel in analytical and screening laboratories.

The plate is also called a microplate, microtiter plate, microwell plate or multiwell plate: the terms are interchangeable. “Microtiter” began as a trade name in the 1950s and stuck. Wells are addressed by row letter (A–H) and column number (1–12), giving unique coordinates such as C7 that anchor every plate map.

Purpose & benefits

Three properties make the 96-well plate indispensable:

• Parallelism. Ninety-six wells mean 96 samples, a full standard curve, or many replicates processed at once, with the same reagents, timing and conditions across all wells.
• Standardization. A fixed footprint lets the plate drop into readers, washers, shakers and robots from any manufacturer.
• Miniaturization. Microlitre volumes cut reagent cost and sample use versus test tubes, while remaining easy to handle by multichannel pipette.

The standard format

The 96-well layout is defined by the ANSI/SLAS microplate standards (formerly SBS standards). They fix the outer footprint and well spacing so labware is interchangeable across the industry. The 96-well is the reference format from which the 384- and 1536-well plates are derived by halving the pitch.

Dimensions & size

Key measurements of a standard (non-deep) 96-well plate:

For plate height by type, the flat-bottom well profile and a 96-vs-384 comparison, see the dedicated 96-well plate dimensions page.

Well volume: how much a well holds

A standard flat-bottom well has a working volume of 100–200 µL and a maximum of about 360 µL. Working volume is the amount used in practice, enough to cover the well base and give a clean optical path, while the maximum is the physical brim capacity, rarely filled to avoid spillage and cross-contamination. A deep-well 96 plate holds far more, typically 1–2 mL per well, for storage and extraction. Compare all formats on the surface area & volume chart.

Well-bottom shapes

The shape of the well base is chosen to match the assay and readout:

Colour & optical options

Plate colour controls stray light and is matched to the detection method:

• Clear: for absorbance (colorimetric) reads and brightfield imaging.
• Black: absorbs light to minimise background and cross-talk in fluorescence.
• White: reflects light to maximise signal in luminescence assays.
• Black or white with a clear bottom: combines low-background walls with an optical base for bottom-reading and live-cell imaging.

Common 96-well variants

Beyond bottom shape and colour, the 96-well format ships in several purpose-built variants:

• Strip-well plates: 8- or 12-well breakable strips in a frame, the standard choice for ELISA when you want to run only part of a plate.
• Half-area plates: narrower wells that halve the volume (~25–100 µL) to save reagent in absorbance and fluorescence assays.
• PCR plates (skirted, semi-skirted, non-skirted): thin-wall 96-well plates that fit a thermal cycler block; the skirt style is chosen for the instrument and sealing method.
• Ultra-low-attachment plates: a coating that prevents adhesion, used to grow spheroids and organoids in suspension.

Materials

Most plates are polystyrene: transparent, inexpensive and readily tissue-culture (TC) treated so adherent cells attach and spread. Polypropylene plates resist solvents (e.g. DMSO) and freeze–thaw, making them the choice for compound storage and PCR. For UV measurements, ordinary polystyrene absorbs UV, so UV-transparent plastics (such as cyclic-olefin) or quartz plates are used instead. Plates are supplied sterile (for cell culture) or non-sterile (for biochemistry).

Applications

The format underpins most parallel laboratory methods:

See what well plates are used for for a fuller breakdown.

Instruments & accessories

A 96-well plate is read and handled by a microplate reader (or plate spectrophotometer), and processed with plate washers, shakers/incubators, centrifuges and robotic liquid handlers, all built to the standard footprint. Common accessories include lids, adhesive and heat-seal films for storage and PCR, and sealing mats.

Common brands

Widely used 96-well plates include Corning (and its Costar line), Greiner Bio-One, Thermo Fisher (Nunc), Eppendorf, Sarstedt and Azenta. All share the standard footprint, so the templates and tools on this site apply regardless of brand.

How to choose a 96-well plate

How to seed cells in a 96-well plate

For adherent cell assays, a typical seeding density is roughly 1–5 × 10⁴ cells per well in 100–200 µL of medium, adjusted for cell type, growth rate and assay length so wells reach the target confluency at read-out. Dispense a master cell suspension with a multichannel pipette, keep the plate level, and leave outer wells as medium-only blanks if edge effects are a concern.

Cost

Standard 96-well plates are inexpensive, typically a few dollars each, less in bulk. Specialty plates (TC-treated, black/white, sterile, low-binding or UV-transparent) cost more. The templates and plate designer on this site are free.

Frequently asked questions