CRISPR cell line development Use Cases with FluidFM®


Accelerate CRISPR gene editing and cell line development.

Transient expression of target proteins with mRNA

Human Dermal Fibroblast injected with GFP mRNA. 70% of injected cells expressed GFP 24h after injection.

Transient mRNA transfection helps to give fundamental insights on protein functions at a single cell level, such as protein expression and intercellular communication.


mRNA transfection using FluidFM nano-injection results in:

  • Higher transfection efficiency

  • Lower cell toxicity

  • Faster expression

Human dermal fibroblast expressing GFP, 24h after injection with FluidFM

Human dermal fibroblast expressing GFP, 24h after injection.

Generation of monoclonal multiple KO clones in <3 weeks

Start from a single cell clone and get your monoclonal cell line within 3 weeks, compared to 10-14 weeks with conventional approaches.  

Injection of multiple gRNAs & Cas9 with FluidFM

Day 1:

Injection of multiple gRNAs & Cas9

Multiple gRNAs are simultaneously delivered with the Cas9 into CHO cells. A fluorescent marker is co-injected to verify successful delivery.


Hands-on time: 30 minutes - from the loading of the FluidFM Nanosyringe until the injection of 50 cells is completed

With FluidFM isolated KO clone

Day 2:

Isolation of the clone candidate

Selected cells are isolated into separate wells. Visual proof of single cell deposition ensures monoclonality.


Hands-on time: 30 minutes - for the preparation of the FluidFM Micropipette and the isolation of 12 injected cells expressing the fluorescent marker

Expansion of KO clones

As of day 3:

Clone expansion and analysis

Of the isolated clones, 90% developed into monoclonal colonies and 50% thereof showed mutations in targeted loci.


Strong reduction in complexity and development time of genetically modified cell lines starting from few nano-injected cells

Direct intra-cellular delivery & volume measurement


Co-delivery of two plasmids

pEGFP-UHRF1 and pmCherry-TRIM21 (2 plasmids) co-injected into adherent CHO-K1 cells. The nuclear expression of the human UHRF1 protein is observed (3h post injection), as well as the cytoplasmic expression of the human TRIM21 protein.

Intra-nuclear CRISPR delivery

Mouse primary hepatocyte injected with CRISPR-Cas9 RNP complexes.



Volume measurement

Measured volumes of fluorescent CRISPR-Cas9 complexes after injection into mouse primary hepatocytes.

Successful transfection of primary neurons with plasmids

With our system, even primary neurons can be transfected successfully and highly efficiently. The FluidFM nanosyringe directly delivers the complexes into primary neurons - gently and autonomously.


Conventional delivery methods applied to neurons are often inefficient, expensive and toxic to the neuron.

Injected neuron. Courtesy Sen Yan, Jinan University.

GFP expression in neurons 24h after nano-injection of plasmids in a mouse primary neuron. Courtesy of Jinan University Guangzhou, China.

Do you want to discuss your experiment with our FluidFM experts?

FluidFM enables novel experiments in CRISPR cell line development - from easy multiplexing to fast cell line development in <3 weeks.