Intracellular　delivery　of　materials　with　nanopipettes　has　become　a　critical　method　in　gene　editing　and　cell-based　therapy　because　of　its　nanometer-scaled　features　capable　of　penetrating　the　cell　membrane　and　targeting　specific　subcellular　compartments-directing　materials　to　the　endoplasmic　reticulum,　nucleus,　or　mitochondria,　for　example.　The　geometrical　parameters　of　nanopipettes　directly　affect　delivery　efficiency.　However,　the　current　methods　for　fabricating　nanopipettes　have　the　disadvantages　of　poor　controllability,　high　cost,　and　difficult　operation.　To　overcome　these　issues,　we　propose　a　real-time　visualization　method　for　fabricating　nanopipettes　based　on　pressured　electrolyte　chamber　based　wet　etching　to　precisely　determine　the　tip　size　during　the　fabrication　process.　A　standard　curve　is　plotted　to　provide　a　direction　for　fabrication,　and　tip　inner　diameters　smaller　than　10　nm　can　be　controllably　achieved.　Furthermore,　the　intranuclear　injection　of　proteins　to　living　single　cells　(diameter　＜　30　mu　m)　with　a　high　spatial　resolution　is　realized.　And　single-cell　transfection　through　the　intracellular　delivery　of　plasmid　based　on　a　self-built　living　single-cell　workstation　is　completed.　This　technique　is　expected　to　be　used　in　the　treatment　of　diseases,　for　high-resolution　localization　of　organelles　in　living　single　cells　without　fluorescent　labeling,　and　for　subcellular　omics　analysis　by　mass　spectrometry.