Clarity in cell line development

VIPS

VIPS™ (Verified in-situ plate seeding) is a unique All-in-One instrument which does the work of two instruments: first of all it performs the single cell seeding into 96 well plates and verifies the success of this process by imaging the nanolitre (nL) volume droplets, and secondly it performs the whole well imaging of the single cell for the assurance of clonality requirements

VIPS uses very gentle, low pressure dispensing of cells in nL droplets into the bottom of dry wells of 96 well microtiter plates. Z-stack imaging confirms the presence of a cell in the droplet, and upon confirmation of a cell, then the well is immediately filled with media. Cell droplets continue to be added to a well until a cell is detected and this ensures high seeding efficiencies.

VIPS Seeding Efficiency – at the end of the seeding run the plate map shows which wells have single cells.

  • Green wells = single cells
  • Red wells = >1 cell
  • Grey wells = no cells

This particular example shows 87% seeding efficiency

Key Features

 

  1. High seeding efficiency – seeding efficiencies of over 85% can be achieved with optimization and depending on cell type
  2. VIPS uses much lower pressure than FACS; VIPS dispensing pressure is less than 1 psi which means that cell survival rates are high
  3. Outgrowth is customer cell line dependent, but the gentle nature of VIPS means cloning efficiency (% of confirmed single cells in the plate which grow into colonies) will be the same or better than the same cell line dispensed by manual limiting dilutions
  4. Single cells are detected in wells, compared with single cell printers which image cells “on the way” to the well
  • Z-stack imaging of cell in nL droplets at the bottom of a dry well (before any media is added)
  • Droplet image of the single cell arriving in the plate is included in the Clonality Report for providing to the regulator
  • Performs the Day 0 whole well image when media has been added to the well
  • Takes only approx. 10 minutes to process an entire 96 well plate (with no further separate imaging time required compared with other single cell printers)
  • Eliminates traditional problems of well edges in whole well imaging because the droplet and cell detection are in the central portion of the well

Optional Features

 

  • Daily whole well imaging to monitor growth and expansion of the clones into colonies (alternatively this can be carried out on the Cell Metric outside the hood)
  • Fluorescence (Red and Green) primarily for validation of the system

Benefits

  • Eliminates need for two separate instruments (a single cell printer and a whole well imager)
  • Only a single round of cloning is required
  • Higher efficiency of seeding and outgrowth means fewer plates needed per project
  • Cell reservoir can be cleaned and reused, and is compatible with wide range of cells – no need for costly proprietary consumables
  • Automatic identification and confirmation of single cells in wells at time of seeding speeds up processes as only clonal wells need to be tracked
  • Data connectivity with existing Cell Metric for daily imaging of cell division and outgrowth
  • Unlike FACS or novel Optofluidic systems, VIPS does not need a dedicated specialist to run the system and anyone in the group should be able to use it after a short training
  • The ‘cell in the droplet’ design and detection provides the highest confidence in clonality
  • Low pressure cell dispensing and no requirement for centrifugation means delicate and fragile cells can be isolated and grown successfully
  • Very low incidence of ghost wells/false negatives (only 0.06% per Janssen Case Study) results in higher levels of assurance for regulators
  • Optional process quality control using fluorescence available for validation experiments
  • The industry-standard Clonality Report is now extended so that it links whole well colony outgrowth image all the way back to image of single cell in dispensed droplet